With the exception of bacteria, living cells contain most of their DNA inside a structure called the nucleus. The membranes of the nucleus form a protective wall around the DNA, while pores within this wall act as entry check-points, controlling what can and cannot get inside. Maintaining the structure of this wall is critical for cell survival. Problems can occur if the nuclear wall or its pores become disrupted, as in the case of cancer and neurodegenerative diseases. Thankfully cells have developed a protective surveillance system that can rapidly identify and repair any damage made to the nuclear wall. However, how this damage is found and what activates its repair is poorly understood.

Now, Thaller et al. have investigated two key proteins that they suspected were involved in the surveillance of the nuclear border in budding yeast: Chm7 and Heh1. Chm7 is part of a complex group of proteins that can cut and sculpt the shape of membranes, while Heh1 is normally embedded on the inside of the nucleus. Thaller et al. discovered that, when the nuclear wall is disrupted, Heh1 recruits Chm7 to the site of damage and activates it. Once activated Chm7 can repair the damage to the nuclear wall, by sealing over defective nuclear pores and closing gaps caused by breakages.

Thaller et al. showed that the transport system that normally moves molecules into and out of the nucleus also imports Heh1 and actively excludes Chm7, physically segregating them to opposite sides of the nuclear border. If the nuclear wall becomes damaged this leads to the local meeting of Heh1 and Chm7 at these sites. Heh1 will then activate the membrane shaping mechanisms of Chm7, rapidly repairing the nuclear border in response to the damage.

It is possible cell structures other than the nucleus use a similar surveillance system to protect their borders. Manipulating the border surveillance system of the nucleus could be used to treat the detrimental impacts of damage caused to the nuclear wall by disease.

The molecular machinery that biochemically segregates the nucleus and the cytoplasm has been extensively investigated. The foundational components of this selective barrier include the double-membrane nuclear envelope with embedded nuclear pore complexes (NPCs). NPCs impose a ‘soft’ diffusion barrier to macromolecules larger than ~40 kD (Popken et al., 2015; Timney et al., 2016) while providing binding sites for the rapid and selective transport of signal-bearing (nuclear localization and nuclear export signals; NLSs and NESs) macromolecules, which are ferried through the NPC by shuttling nuclear transport receptors (NTRs; a.k.a. karyopherins/importins/exportins; Schmidt and Görlich, 2016). Directionality and energy for NTR-selective transport is imparted by the spatial segregation of the Ran-GTPase whose nuclear GTP-bound form destabilizes and stabilizes import and export complexes, respectively (Floch et al., 2014).

Interestingly, the robustness of the nuclear envelope barrier has been shown to be compromised in several different contexts, including in diverse human diseases (Hatch and Hetzer, 2014; Lusk and King, 2017). For example, there is an emerging body of work linking the function of NTRs and NPCs with neurodegenerative diseases like ALS and FTD (Nousiainen et al., 2008; Freibaum et al., 2015; Jovičić et al., 2015; Kaneb et al., 2015; Zhang et al., 2015; Kim and Taylor, 2017; Shi et al., 2017). These studies, coupled to the observations of age-related declines in NPC function in both post-mitotic multicellular systems (D'Angelo et al., 2009; Savas et al., 2012; Toyama et al., 2013) and also in replicative aging models like budding yeast (Janssens et al., 2015; Lord et al., 2015), support a theme in which the function of the nuclear envelope could be mitigatory of age-related disease progression (Schreiber and Kennedy, 2013; Jevtić et al., 2014; Serebryannyy and Misteli, 2018). Of similar interest, the hallmark cellular pathophysiology of early-onset dystonia are nuclear envelope herniations (Goodchild et al., 2005) that emanate from NPC-like structures (Laudermilch et al., 2016). As analogous herniations have been observed in many genetic backgrounds associated with defects in NPC biogenesis in yeast over several decades (Thaller and Lusk, 2018), this has contributed to the idea that the herniations might be the result of either defective NPC assembly events (Scarcelli et al., 2007; Onischenko et al., 2017; Zhang et al., 2018) and/or the triggering of a NPC (Wente and Blobel, 1993) or NPC assembly quality control pathway (Webster et al., 2014; Webster et al., 2016). The latter could depend on the function of the endosomal sorting complexes required for transport (ESCRT), a membrane scission machinery that has been proposed to seal-off malforming NPCs (Webster et al., 2016).

That there could be mechanisms to surveil the assembly of NPCs makes considerable sense as there are hundreds of NPCs, each containing hundreds of nucleoporins/nups (Kosinski et al., 2016; Kim et al., 2018), that are assembled during interphase in mammalian cells (Maul et al., 1972; Doucet et al., 2010; Dultz and Ellenberg, 2010). There are approximately 100 NPCs formed during a budding yeast cell cycle, which includes a closed mitosis (Winey et al., 1997). As interphase NPC assembly likely occurs through an inside-out evagination of the inner nuclear membrane (INM) followed by membrane fusion with the outer nuclear membrane (ONM) (Otsuka et al., 2016), holes are constantly being formed in the nuclear envelope. Without mechanisms to surveil this process, de novo NPC biogenesis might pose a threat to nuclear-cytoplasmic compartmentalization (Webster et al., 2014). Consistent with this idea, malformed or damaged NPCs are not passed on to daughter cells in budding yeast (Colombi et al., 2013; Makio et al., 2013; Webster et al., 2014). Further, deletion of the ESCRT machinery in the context of genetic backgrounds where nuclear envelope herniations have been observed for example nup116Δ (Wente and Blobel, 1993) or apq12Δ (Scarcelli et al., 2007) cells require a nuclear envelope-specific ESCRT, Chm7 (the orthologue of mammalian CHMP7), for viability (Bauer et al., 2015; Webster et al., 2016). While we have previously proposed that a biochemical signature of malforming NPCs is surveilled by integral inner nuclear membrane proteins of the Lap2-emerin-MAN1 (LEM) domain family, specifically Heh2, it remains to be formally established what the signal that leads to ESCRT recruitment to the nuclear envelope actually comprises (Webster et al., 2014).

Evidence that the ESCRT machinery acts at holes in the nuclear envelope is further exemplified by their critical role in performing annular fusion events during the final stages of nuclear envelope reformation at the end of mitosis in mammalian cells (Olmos et al., 2015; Olmos et al., 2016; Vietri et al., 2015; Gu et al., 2017; Ventimiglia et al., 2018). Moreover, ESCRTs are also required for the efficient repair of nuclear ruptures that arise during the migration of cells through tight constrictions (Denais et al., 2016; Raab et al., 2016). And, it is most likely that they also act to repair nuclear envelope ruptures that are induced by intracellular mechanical stresses from either the actin cytoskeleton (Hatch and Hetzer, 2016; Robijns et al., 2016), or from those observed during telomere crisis (Maciejowski et al., 2015). Lastly, recent work also suggests a role for ESCRTs in the context of turning over NPCs in quiescent cells (Toyama et al., 2019). It remains an open question, however, whether the mechanisms that repair nuclear ruptures, seal the nuclear envelope at the end of mitosis, and protect against defective NPC assembly respond to an identical upstream signal and proceed through the same membrane-sealing mechanism.

Clues to what might constitute the upstream signal that leads to nuclear envelope-recruitment of ESCRTs could be drawn from other contexts where ESCRTs protect membrane compartments including endolysosomes (Radulovic et al., 2018; Skowyra et al., 2018) and the plasma membrane (Jimenez et al., 2014; Scheffer et al., 2014; Gong et al., 2017). In both of these cases, there is evidence to suggest that the local release of Ca²⁺ is a trigger for ESCRT recruitment, through (at least at the plasma membrane) a Ca²⁺ binding protein, ALG-2 (Jimenez et al., 2014; Gong et al., 2017). Whether Ca²⁺ plays a role at the nuclear envelope remains unaddressed. More generally, there are two, often redundant, recruitment mechanisms seeded by either an ESCRT-I, II complex and/or ESCRT-II and ALIX (Bro1 in yeast) that bind and activate ESCRT-III subunit polymerization (Wemmer et al., 2011; Henne et al., 2012; Tang et al., 2015; Tang et al., 2016; Christ et al., 2016) on specific membranes throughout the cell (reviewed in Schöneberg et al., 2017; McCullough et al., 2018).

ESCRT-III polymers predominantly made up of the most abundant ESCRT-III (Snf7/CHMP4B) scaffold negative but also in at least one case, positive membrane curvature (McCullough et al., 2015), and directly contribute to membrane scission (Adell et al., 2014; Adell et al., 2017; Schöneberg et al., 2018). The AAA +ATPase Vps4 disassembles ESCRT-III filaments by directly interacting with MIM (MIT interacting motif) domains present on a subset of ESCRT-III subunits (Obita et al., 2007; Stuchell-Brereton et al., 2007; Agromayor et al., 2009; Xiao et al., 2009; Han et al., 2015) by threading the ESCRT-III filaments through the central cavity of a hexameric ring (Yang et al., 2015; Han et al., 2017; Monroe et al., 2017; Su et al., 2017). It is likely that ESCRT-III disassembly by Vps4 directly contributes force to promote membrane scission (Schöneberg et al., 2018). Whether the membrane scission reaction is different in distinct subcellular contexts like at the nuclear envelope remains to be understood.

Consistent with the idea that there might be unique ESCRT membrane remodeling mechanisms at play in distinct compartments, a step-wise recruitment and activation mechanism requiring the ESCRT-II Vps25 and the ESCRT-III Vps20 is thought to be required at budding yeast endosomes (Saksena et al., 2009; Teis et al., 2010; Tang et al., 2015; Tang et al., 2016), but both of these proteins are absent from the genetic and biochemical analyses of the nuclear envelope arm of the ESCRT pathway (Webster et al., 2014; Webster et al., 2016). These data suggest that other proteins likely contribute to ESCRT-III activation at the nuclear envelope. Key candidates are Chm7 and the inner nuclear membrane (INM) proteins, Heh1/Src1 (orthologue of LEM2) and Heh2 (orthologue of MAN1 or other LEM-domain proteins). These proteins have collectively been shown to interact biochemically and genetically with Snf7 (Webster et al., 2014; Webster et al., 2016) and Heh1 is required for the focal accumulation of Chm7 at the nuclear envelope in genetic backgrounds where NPC assembly is inhibited (Webster et al., 2016). Remarkably, the interactions between Heh1 and Chm7 are well conserved in both fission yeast (Gu et al., 2017) but also in mammalian cells, where LEM2 is required to recruit CHMP7 to the reforming nuclear envelope at the end of mitosis (Gu et al., 2017). It remains to be understood, however, whether LEM proteins (or Chm7) directly contribute to ESCRT-III activation at the nuclear envelope or whether additional proteins are involved.

Heh1 and Heh2 contain an N-terminal helix-extension-helix (heh) motif (the LEM domain), followed by an INM targeting sequence that (at least in the case of Heh2 but likely also Heh1 [King et al., 2006; Lokareddy et al., 2015]) includes an NLS and a ~ 200 amino acid unstructured region that are both required for INM targeting (Meinema et al., 2011). They both also contain a second nuclear-oriented domain, which likely folds into a winged helix (WH) (also called MAN1/Src1-C-terminal homology domain or MSC; Caputo et al., 2006); this domain is also well conserved through evolution (Mans et al., 2004; Mekhail et al., 2008). The LEM domain proteins as a family have been ascribed diverse roles in gene expression either through binding to transcription factors, BAF, or the lamins (Barton et al., 2015). While yeasts lack BAF and lamins, the LEM domain proteins nonetheless directly interface with chromatin (Grund et al., 2008; Barton et al., 2015), contribute to rDNA repeat stability (Mekhail et al., 2008) and the mechanical robustness of the nucleus (Schreiner et al., 2015). This latter function is likely revealed by the observation in many diverse yeast species that loss of Heh1 leads to nuclear envelope disruption (Yewdell et al., 2011; Gonzalez et al., 2012; Yam et al., 2013), however, it might also reflect Heh1’s role in recruiting ESCRTs to the nuclear envelope. Thus, the relationship between how the LEM proteins contribute to nuclear integrity and ESCRT-mediated surveillance remains to be clearly defined.

In the following, we further explore the molecular determinants of Chm7 recruitment to the budding yeast nuclear envelope by Heh1. We determine that the spatial segregation of Chm7 and Heh1 on either side of the nuclear envelope is driven by NTRs and the robustness of the nuclear transport system. Perturbations of this system, or the exposure of Heh1 to the cytosol leads to the local recruitment and Heh1 WH-dependent activation of Chm7. At sites of Chm7 hyperactivation, we observe remarkable alterations to nuclear envelope morphology including nuclear envelope herniations and intranuclear INM invaginations suggesting a role for membrane expansion and remodeling during nuclear envelope repair.

While there has been considerable focus over the last few decades on mechanisms that control the targeting of proteins and lipids to distinct intracellular compartments, it is equally important to understand the protective mechanisms that maintain this compartmentalization in the face of challenges to membrane integrity and/or the specific biochemical identity of organelles. Here, we further explore the mechanism of ESCRT surveillance of the nuclear envelope. We interpret our data in a model where the nuclear envelope is surveilled by two principle components, the ESCRT Chm7 and the integral INM protein, Heh1. This surveillance system appears to be set up to respond directly to perturbations in the nuclear envelope barrier in a way that we suggest is agnostic as to whether the perturbation is a result of defectively formed NPCs or a mechanical (or other) disruption of the nuclear membranes.

The rationale behind this assertion is that the nuclear envelope surveillance system is itself directly established by a functioning nuclear transport system, which physically segregates Chm7 and Heh1 on either side of the nuclear envelope. For example, prior work has shown that Heh1 requires the function of the NTRs Kap-α and Kap-β1 in addition to the Ran-GTPase in order to be actively targeted to the INM through NPCs (King et al., 2006). Here, we establish that Chm7, while small enough to passively leak through the NPC diffusion barrier, is actively exported by the major export NTR, Xpo1 (Figure 1). Therefore, any perturbations that impact active nuclear transport or the diffusion barrier across the nuclear envelope would lead to increased Chm7 diffusion into the nucleus and/or a deficit in its nuclear export increasing the likelihood that it meets Heh1. While similar perturbations could also lead to Heh1 mistargeting and/or its diffusion into the ONM, we suspect that this would be kinetically slower than Chm7 diffusion into the nucleus as Heh1 is bound to chromatin at the INM (Grund et al., 2008; Mekhail et al., 2008; Gonzalez et al., 2012; Yam et al., 2013; Barton et al., 2015; Schreiner et al., 2015). Indeed, it is probable that Heh1 functions in two major roles with respect to nuclear integrity: first, it provides mechanical stability to the nucleus by binding chromatin, and second, provides a binding site for Chm7 through its C-terminal WH domain. These two inter-related roles could also help to explain why both the N- and C-terminal domains of Heh1 are required to maintain viability of apq12Δ cells (Figure 2C), and why HEH1 is generally more essential in budding and fission yeasts compared to CHM7. This likely holds true in mammalian models as well as LEM2 is essential whereas CHMP7 is dispensable for viability (in cell culture) (Hart et al., 2015).

Once a perturbation occurs to the nuclear envelope barrier through defects in NPC assembly, loss of function of NPCs, or mechanical disruption of the nuclear membranes, Chm7 and Heh1 are able to come together. While we have so far been unsuccessful in reconstituting a direct biochemical interaction between Chm7 and Heh1 (although others have between LEM2 and CHMP7 [Gu et al., 2017]), we have nonetheless provided evidence that this interaction likely leads to Chm7 activation, thus tightly coupling recruitment and activation (Figure 4). Our data support a model in which the C-terminal WH domain of Heh1 in addition to a membrane anchor are necessary and sufficient for this activation event. As our prior work (Webster et al., 2016) and that from Olmos et al. (2016), support that it is the N-terminal ESCRT-II-like domain of Chm7 (which, interestingly, is also predicted to be made up of tandem WH domains [Horii et al., 2006; Bauer et al., 2015; Figure 1A]) that is necessary for recruitment to the nuclear envelope, it seems likely that the binding between the Heh1-WH domain and the N-terminus of Chm7 could trigger activation by removing some form of autoinhibition, which is a common theme among ESCRT-III proteins (Zamborlini et al., 2006; Shim et al., 2007; Lata et al., 2008; Bajorek et al., 2009; Henne et al., 2012; Tang et al., 2015; Tang et al., 2016) that ensures polymerization in the correct compartment. Clearly, the precise molecular mechanism of Chm7 activation by Heh1 will require structural insight, which is no doubt on the horizon.

Even with structural information, there are many additional factors that need to incorporated into a nuclear envelope surveillance mechanism. For example, future work must directly address how Heh2 fits into this pathway. While Heh2 has a similar domain architecture as Heh1 including a WH domain (King et al., 2006), it does not appear to impact Chm7 activation within the FETA assay (Figure 5—figure supplement 1D), nor was it detected in any of the affinity purifications (Figure 3). This was surprising, as we had previously established that the N-terminus of Heh2 (that lacks the WH domain), can directly bind to both Chm7 and Snf7, at least in their ‘open’ forms (Webster et al., 2016). Moreover, we reported physical interactions between Chm7 and Snf7 with Heh2 in vivo (Webster et al., 2014; Webster et al., 2016). While there are many possible explanations for these results, we favor models that consider Heh2 in a regulatory role that might modulate Chm7 and/or Snf7 function at the nuclear envelope, although this remains speculative and awaits direct experimentation.

Additional downstream (of Chm7 and Heh1) components also need to be figured into any nuclear envelope surveillance mechanism including Snf7, but also Did2, Vps24 and Vps2 (Figure 3B,C). That these ESCRT-IIIs impact the recruitment of Vps4 to the nuclear envelope (Figure 3E) raises the possibility that the sequence of their recruitment might be similar to that found at ILVs during MVB formation, which begins with Snf7 and ends with Did2, Vps24, and Vps2 (Babst et al., 2002; Teis et al., 2008), the latter of which play important roles in Vps4 recruitment and ATPase activation required for membrane scission (Azmi et al., 2008; Shestakova et al., 2010; Schöneberg et al., 2018). Interestingly, however, while we observe Vps4 recruitment to sites of chm7_OPEN at the INM (Figure 3E), we do not have any explicit evidence for membrane scission per se. Indeed, the INM evaginations and nuclear envelope herniations appear to be stabilized by ~45 nm diameter necks (Figures 6, 7 and 8B). That these necks have similar diameters to those observed at the plasma membrane or endosomes when HIV budding (von Schwedler et al., 2003; Morita et al., 2011; Cashikar et al., 2014; Jackson et al., 2017) or ILV formation (Adell et al., 2014; Buono et al., 2017; Frankel et al., 2017; Wenzel et al., 2018), respectively, are stalled, is suggestive that they are stabilized by a similar ESCRT-III-like polymer. We suggest that the most likely explanation is that Chm7 itself might play a unique role in a nuclear membrane-scission step, perhaps through its potential MIM1 domain that might directly recruit Vps4 but is absent from the chm7_OPEN construct. Such a conclusion is supported by the presence of similar INM evaginations and nuclear envelope herniations observed in the presence of full length Chm7 but lacking Vps4 (Figure 7).

Any scission mechanism that requires a direct interaction between the MIM1 domain of Chm7 and Vps4 will also need to consider the overlap of this sequence with NES2 (Figure 1). This clearly predicts a competition between Xpo1 and Vps4 for Chm7, which provides a potential entryway for an additional level of regulation for any polymer formation or scission reaction by this NTR (that would also be modulated by Ran-GTP). Indeed, NTRs might regulate this pathway at the level of Heh1 as well. Heh1 is synthesized and inserted into ER membranes so is exposed to the cytosol and would thus be capable of binding to Chm7 before it is targeted to the INM. Such an assumption is supported by the observation that the exposure of the WH domain of Heh1 is sufficient to recruit Chm7 to ER membranes (Figure 2). Does NTR-binding to the NLSs in the N-terminus of Heh1 inhibit Chm7 recruitment in this compartment? Answering this question will also be relevant in mammalian systems where LEM2 and CHMP7 are found in the same compartment during mitotic nuclear envelope breakdown suggesting the need for mechanisms to prevent their binding in this phase of the cell cycle. Interestingly, recent work is suggesting that proteins like Lgd/CC2D1B help control the spatiotemporal timing of CHMP4B and CHMP7 activity in mammalian cells (Ventimiglia et al., 2018), supporting the existence of regulatory mechanisms of this pathway.

Lgd/CC2D18 acts during nuclear envelope reformation when ESCRTs play critical roles in sealing the nuclear envelope at the end of mitosis, often at sites where spindle microtubules perforate the nascent nuclear envelope (Olmos et al., 2015; Olmos et al., 2016; Vietri et al., 2015). It will be interesting to understand the similarities and differences between how ESCRTs function at the end of mitosis (in an open mitosis) versus events that are triggered upon acute disruption of nuclear envelope integrity or NPC function during interphase. For example, two of the more interesting observations of the EM tomographic analyses are the generation of intranuclear membrane invaginations and the proximity of ER sheets and vesicles at the ONM sites of nuclear envelope herniations (Figure 6, Figure 6—figure supplement 1, Figure 7). Both of these observations suggest that nuclear envelope surveillance might be coupled to the delivery of new membrane at sites of rupture. It seems plausible that sealing holes in the nuclear envelope (particularly those larger than a nuclear pore) would require the local delivery of membranes. Similarly, NPC quality control mechanisms that suggest the sealing of defective NPCs would likely require some form of expansion of the pore membrane, as has been proposed (Wente and Blobel, 1993). Whether such membrane is derived from new synthesis or from the mobilization of existing stores remains to be explored. Of note, recent work supports that even the INM may be metabolically active and have the capacity to generate new lipid locally (Romanauska and Köhler, 2018). Further, close examination of sites of chm7_OPEN accumulation reveals that the morphology of the INM extensions are both sheet-like and tubular in nature and resemble the ‘normal’ continuities between the ONM and the broader ER network (West et al., 2011); together these data suggest that ER-shaping proteins and lipid synthesis pathways might also play an important role in contributing to nuclear envelope sealing - topics for the future.

Both ER remodeling proteins and lipid synthesis pathways are required for de novo NPC biogenesis (Schneiter et al., 1996; Dawson et al., 2009; Hodge et al., 2010). As NPC biogenesis proceeds through an INM evagination step, we also must consider whether the INM evaginations observed in the electron tomograms reflect a function for Chm7 in this process. However, while this remains a compelling hypothesis, our data do not yet support this idea. For example, the herniations observed in the context of inhibiting NPC assembly are morphologically distinct from those found in the chm7_OPEN or vps4Δpom152Δ scenarios with wider necks that contain nups (Figure 8A,B). Further, they arise in the absence of CHM7 and are thus more likely to be formed due to a defect in NPC biogenesis (perhaps because of an inhibition of INM-ONM fusion), or, through the triggering of a NPC assembly surveillance mechanism (Webster et al., 2016; Thaller and Lusk, 2018). Should Chm7 be a critical component of the latter mechanism, it would be predicted that the nuclear envelope herniations associated with nups would be unsealed without CHM7. Indeed, that we observe often-dramatic openings in the nuclear envelope in apq12Δchm7Δ cells (Figure 8D and Figure 8—figure supplement 2), suggest that the herniations themselves might be prone to rupture with Chm7 required for their repair. It follows then that like in mammalian cells where much larger nuclear envelope herniations are precursors to nuclear rupture (De Vos et al., 2011; Vargas et al., 2012; Hatch et al., 2013; Denais et al., 2016; Hatch and Hetzer, 2016; Raab et al., 2016), these smaller NPC-assembly associated herniations might also impact nuclear envelope integrity through mechanisms that remain to be fully defined. In either case, it reinforces the concept that the assembly of NPCs can be perilous, and it will be important to consider this possibility when interpreting the underlying pathology of human diseases that are associated with defects in NPC function or assembly, for example, DYT1 early-onset dystonia (Laudermilch et al., 2016; Pappas et al., 2018) or Steroid Resistant Nephrotic Syndrome (Miyake et al., 2015; Braun et al., 2016; Braun et al., 2018).

Lastly, while we acknowledge then that the INM evaginations that we observe in cases of Chm7 hyperactivation might not necessarily be a physiological event in a nuclear envelope sealing process, it remains tempting to speculate that they might be in the context of proposed mechanisms of nuclear egress be it Mega-RNPs (Speese et al., 2012; Jokhi et al., 2013), viruses (Lee et al., 2012; Lee et al., 2016; Arii et al., 2018), or nucleophagy (Roberts et al., 2003; Dou et al., 2015; Mochida et al., 2015; Mostofa et al., 2018) that so-far remain obscure but would nonetheless require a membrane scission step. Interestingly, recent work suggests that herpes virus nuclear egress requires ESCRTs (Arii et al., 2018), including a role in controlling INM extensions into the nucleus; such intranuclear membrane might also be relevant in cell types that have so-called ‘nucleoplasmic reticulum’ (Malhas et al., 2011). The biogenesis and function of nucleoplasmic reticulum remain enigmatic but our observations of intranuclear fenestrated membrane emanating from the INM might suggest a yet-to-be discovered role for Chm7 and Heh1 in forming such structures as well.

All data generated and analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figure 3.

1. 1. Adell MA
   2. Vogel GF
   3. Pakdel M
   4. Müller M
   5. Lindner H
   6. Hess MW
   7. Teis D

   (2014) Coordinated binding of Vps4 to ESCRT-III drives membrane neck constriction during MVB vesicle formation

   The Journal of Cell Biology 205:33–49.

   https://doi.org/10.1083/jcb.201310114

   • PubMed
   • Google Scholar
2. 1. Adell MAY
   2. Migliano SM
   3. Upadhyayula S
   4. Bykov YS
   5. Sprenger S
   6. Pakdel M
   7. Vogel GF
   8. Jih G
   9. Skillern W
   10. Behrouzi R
   11. Babst M
   12. Schmidt O
   13. Hess MW
   14. Briggs JA
   15. Kirchhausen T
   16. Teis D

   (2017) Recruitment dynamics of ESCRT-III and Vps4 to endosomes and implications for reverse membrane budding

   eLife 6:e31652.

   https://doi.org/10.7554/eLife.31652

   • PubMed
   • Google Scholar
3. 1. Agromayor M
   2. Carlton JG
   3. Phelan JP
   4. Matthews DR
   5. Carlin LM
   6. Ameer-Beg S
   7. Bowers K
   8. Martin-Serrano J

   (2009) Essential role of hIST1 in cytokinesis

   Molecular Biology of the Cell 20:1374–1387.

   https://doi.org/10.1091/mbc.e08-05-0474

   • PubMed
   • Google Scholar
4. Book

   1. Amberg DC
   2. Burke D
   3. Strathern JN

   (2005)

   Methods in Yeast Genetics: a Cold Spring Harbor Laboratory Course Manual

   Cold Spring Harbor Laboratory Press.

   • Google Scholar
5. 1. Arii J
   2. Watanabe M
   3. Maeda F
   4. Tokai-Nishizumi N
   5. Chihara T
   6. Miura M
   7. Maruzuru Y
   8. Koyanagi N
   9. Kato A
   10. Kawaguchi Y

   (2018) ESCRT-III mediates budding across the inner nuclear membrane and regulates its integrity

   Nature Communications 9:3379.

   https://doi.org/10.1038/s41467-018-05889-9

   • PubMed
   • Google Scholar
6. 1. Azmi IF
   2. Davies BA
   3. Xiao J
   4. Babst M
   5. Xu Z
   6. Katzmann DJ

   (2008) ESCRT-III family members stimulate Vps4 ATPase activity directly or via Vta1

   Developmental Cell 14:50–61.

   https://doi.org/10.1016/j.devcel.2007.10.021

   • PubMed
   • Google Scholar
7. 1. Babst M
   2. Katzmann DJ
   3. Estepa-Sabal EJ
   4. Meerloo T
   5. Emr SD

   (2002) Escrt-III: an endosome-associated heterooligomeric protein complex required for mvb sorting

   Developmental Cell 3:271–282.

   https://doi.org/10.1016/s1534-5807(02)00220-4

   • PubMed
   • Google Scholar
8. 1. Bajorek M
   2. Schubert HL
   3. McCullough J
   4. Langelier C
   5. Eckert DM
   6. Stubblefield WM
   7. Uter NT
   8. Myszka DG
   9. Hill CP
   10. Sundquist WI

   (2009) Structural basis for ESCRT-III protein autoinhibition

   Nature Structural & Molecular Biology 16:754–762.

   https://doi.org/10.1038/nsmb.1621

   • PubMed
   • Google Scholar
9. 1. Barton LJ
   2. Soshnev AA
   3. Geyer PK

   (2015) Networking in the nucleus: a spotlight on LEM-domain proteins

   Current Opinion in Cell Biology 34:1–8.

   https://doi.org/10.1016/j.ceb.2015.03.005

   • PubMed
   • Google Scholar
10. 1. Bauer I
    2. Brune T
    3. Preiss R
    4. Kölling R

    (2015) Evidence for a nonendosomal function of the Saccharomyces cerevisiae ESCRT-III-Like protein Chm7

    Genetics 201:1439–1452.

    https://doi.org/10.1534/genetics.115.178939

    • PubMed
    • Google Scholar
11. 1. Braun DA
    2. Sadowski CE
    3. Kohl S
    4. Lovric S
    5. Astrinidis SA
    6. Pabst WL
    7. Gee HY
    8. Ashraf S
    9. Lawson JA
    10. Shril S
    11. Airik M
    12. Tan W
    13. Schapiro D
    14. Rao J
    15. Choi WI
    16. Hermle T
    17. Kemper MJ
    18. Pohl M
    19. Ozaltin F
    20. Konrad M
    21. Bogdanovic R
    22. Büscher R
    23. Helmchen U
    24. Serdaroglu E
    25. Lifton RP
    26. Antonin W
    27. Hildebrandt F

    (2016) Mutations in nuclear pore genes NUP93, NUP205 and XPO5 cause steroid-resistant nephrotic syndrome

    Nature Genetics 48:457–465.

    https://doi.org/10.1038/ng.3512

    • PubMed
    • Google Scholar
12. 1. Braun DA
    2. Lovric S
    3. Schapiro D
    4. Schneider R
    5. Marquez J
    6. Asif M
    7. Hussain MS
    8. Daga A
    9. Widmeier E
    10. Rao J
    11. Ashraf S
    12. Tan W
    13. Lusk CP
    14. Kolb A
    15. Jobst-Schwan T
    16. Schmidt JM
    17. Hoogstraten CA
    18. Eddy K
    19. Kitzler TM
    20. Shril S
    21. Moawia A
    22. Schrage K
    23. Khayyat AIA
    24. Lawson JA
    25. Gee HY
    26. Warejko JK
    27. Hermle T
    28. Majmundar AJ
    29. Hugo H
    30. Budde B
    31. Motameny S
    32. Altmüller J
    33. Noegel AA
    34. Fathy HM
    35. Gale DP
    36. Waseem SS
    37. Khan A
    38. Kerecuk L
    39. Hashmi S
    40. Mohebbi N
    41. Ettenger R
    42. Serdaroğlu E
    43. Alhasan KA
    44. Hashem M
    45. Goncalves S
    46. Ariceta G
    47. Ubetagoyena M
    48. Antonin W
    49. Baig SM
    50. Alkuraya FS
    51. Shen Q
    52. Xu H
    53. Antignac C
    54. Lifton RP
    55. Mane S
    56. Nürnberg P
    57. Khokha MK
    58. Hildebrandt F

    (2018) Mutations in multiple components of the nuclear pore complex cause nephrotic syndrome

    Journal of Clinical Investigation 128:4313–4328.

    https://doi.org/10.1172/JCI98688

    • PubMed
    • Google Scholar
13. 1. Buono RA
    2. Leier A
    3. Paez-Valencia J
    4. Pennington J
    5. Goodman K
    6. Miller N
    7. Ahlquist P
    8. Marquez-Lago TT
    9. Otegui MS

    (2017) ESCRT-mediated vesicle concatenation in plant endosomes

    The Journal of Cell Biology 216:2167–2177.

    https://doi.org/10.1083/jcb.201612040

    • PubMed
    • Google Scholar
14. 1. Caputo S
    2. Couprie J
    3. Duband-Goulet I
    4. Kondé E
    5. Lin F
    6. Braud S
    7. Gondry M
    8. Gilquin B
    9. Worman HJ
    10. Zinn-Justin S

    (2006) The carboxyl-terminal nucleoplasmic region of MAN1 exhibits a DNA binding winged helix domain

    Journal of Biological Chemistry 281:18208–18215.

    https://doi.org/10.1074/jbc.M601980200

    • PubMed
    • Google Scholar
15. 1. Cashikar AG
    2. Shim S
    3. Roth R
    4. Maldazys MR
    5. Heuser JE
    6. Hanson PI

    (2014) Structure of cellular ESCRT-III spirals and their relationship to HIV budding

    eLife 3:e02184.

    https://doi.org/10.7554/eLife.02184

    • Google Scholar
16. 1. Christ L
    2. Wenzel EM
    3. Liestøl K
    4. Raiborg C
    5. Campsteijn C
    6. Stenmark H

    (2016) ALIX and ESCRT-I/II function as parallel ESCRT-III recruiters in cytokinetic abscission

    The Journal of Cell Biology 212:499–513.

    https://doi.org/10.1083/jcb.201507009

    • PubMed
    • Google Scholar
17. 1. Colombi P
    2. Webster BM
    3. Fröhlich F
    4. Lusk CP

    (2013) The transmission of nuclear pore complexes to daughter cells requires a cytoplasmic pool of Nsp1

    The Journal of Cell Biology 203:215–232.

    https://doi.org/10.1083/jcb.201305115

    • PubMed
    • Google Scholar
18. 1. Craig R
    2. Beavis RC

    (2003) A method for reducing the time required to match protein sequences with tandem mass spectra

    Rapid Communications in Mass Spectrometry 17:2310–2316.

    https://doi.org/10.1002/rcm.1198

    • PubMed
    • Google Scholar
19. 1. Cristea IM
    2. Williams R
    3. Chait BT
    4. Rout MP

    (2005) Fluorescent proteins as proteomic probes

    Molecular & Cellular Proteomics 4:1933–1941.

    https://doi.org/10.1074/mcp.M500227-MCP200

    • PubMed
    • Google Scholar
20. 1. Curwin AJ
    2. Brouwers N
    3. Alonso Y Adell M
    4. Teis D
    5. Turacchio G
    6. Parashuraman S
    7. Ronchi P
    8. Malhotra V

    (2016) ESCRT-III drives the final stages of CUPS maturation for unconventional protein secretion

    eLife 5:e16299.

    https://doi.org/10.7554/eLife.16299

    • PubMed
    • Google Scholar
21. 1. D'Angelo MA
    2. Raices M
    3. Panowski SH
    4. Hetzer MW

    (2009) Age-dependent deterioration of nuclear pore complexes causes a loss of nuclear integrity in postmitotic cells

    Cell 136:284–295.

    https://doi.org/10.1016/j.cell.2008.11.037

    • PubMed
    • Google Scholar
22. 1. Dawson TR
    2. Lazarus MD
    3. Hetzer MW
    4. Wente SR

    (2009) ER membrane-bending proteins are necessary for de novo nuclear pore formation

    The Journal of Cell Biology 184:659–675.

    https://doi.org/10.1083/jcb.200806174

    • PubMed
    • Google Scholar
23. 1. de Chaumont F
    2. Dallongeville S
    3. Chenouard N
    4. Hervé N
    5. Pop S
    6. Provoost T
    7. Meas-Yedid V
    8. Pankajakshan P
    9. Lecomte T
    10. Le Montagner Y
    11. Lagache T
    12. Dufour A
    13. Olivo-Marin JC

    (2012) Icy: an open bioimage informatics platform for extended reproducible research

    Nature Methods 9:690–696.

    https://doi.org/10.1038/nmeth.2075

    • PubMed
    • Google Scholar
24. 1. De Vos WH
    2. Houben F
    3. Kamps M
    4. Malhas A
    5. Verheyen F
    6. Cox J
    7. Manders EM
    8. Verstraeten VL
    9. van Steensel MA
    10. Marcelis CL
    11. van den Wijngaard A
    12. Vaux DJ
    13. Ramaekers FC
    14. Broers JL

    (2011) Repetitive disruptions of the nuclear envelope invoke temporary loss of cellular compartmentalization in laminopathies

    Human Molecular Genetics 20:4175–4186.

    https://doi.org/10.1093/hmg/ddr344

    • PubMed
    • Google Scholar
25. 1. Denais CM
    2. Gilbert RM
    3. Isermann P
    4. McGregor AL
    5. te Lindert M
    6. Weigelin B
    7. Davidson PM
    8. Friedl P
    9. Wolf K
    10. Lammerding J

    (2016) Nuclear envelope rupture and repair during cancer cell migration

    Science 352:353–358.

    https://doi.org/10.1126/science.aad7297

    • PubMed
    • Google Scholar
26. 1. Dimaano C
    2. Jones CB
    3. Hanono A
    4. Curtiss M
    5. Babst M

    (2008) Ist1 regulates Vps4 localization and assembly

    Molecular Biology of the Cell 19:465–474.

    https://doi.org/10.1091/mbc.e07-08-0747

    • PubMed
    • Google Scholar
27. 1. Dou Z
    2. Xu C
    3. Donahue G
    4. Shimi T
    5. Pan JA
    6. Zhu J
    7. Ivanov A
    8. Capell BC
    9. Drake AM
    10. Shah PP
    11. Catanzaro JM
    12. Ricketts MD
    13. Lamark T
    14. Adam SA
    15. Marmorstein R
    16. Zong WX
    17. Johansen T
    18. Goldman RD
    19. Adams PD
    20. Berger SL

    (2015) Autophagy mediates degradation of nuclear lamina

    Nature 527:105–109.

    https://doi.org/10.1038/nature15548

    • PubMed
    • Google Scholar
28. 1. Doucet CM
    2. Talamas JA
    3. Hetzer MW

    (2010) Cell cycle-dependent differences in nuclear pore complex assembly in metazoa

    Cell 141:1030–1041.

    https://doi.org/10.1016/j.cell.2010.04.036

    • PubMed
    • Google Scholar
29. 1. Dultz E
    2. Ellenberg J

    (2010) Live imaging of single nuclear pores reveals unique assembly kinetics and mechanism in interphase

    The Journal of Cell Biology 191:15–22.

    https://doi.org/10.1083/jcb.201007076

    • PubMed
    • Google Scholar
30. 1. Fernandez-Martinez J
    2. Phillips J
    3. Sekedat MD
    4. Diaz-Avalos R
    5. Velazquez-Muriel J
    6. Franke JD
    7. Williams R
    8. Stokes DL
    9. Chait BT
    10. Sali A
    11. Rout MP

    (2012) Structure-function mapping of a heptameric module in the nuclear pore complex

    The Journal of Cell Biology 196:419–434.

    https://doi.org/10.1083/jcb.201109008

    • PubMed
    • Google Scholar
31. 1. Floch AG
    2. Palancade B
    3. Doye V

    (2014) Fifty years of nuclear pores and nucleocytoplasmic transport studies: multiple tools revealing complex rules

    Methods in Cell Biology 122:1–40.

    https://doi.org/10.1016/B978-0-12-417160-2.00001-1

    • PubMed
    • Google Scholar
32. 1. Forster B
    2. Van De Ville D
    3. Berent J
    4. Sage D
    5. Unser M

    (2004) Complex wavelets for extended depth-of-field: a new method for the fusion of multichannel microscopy images

    Microscopy Research and Technique 65:33–42.

    https://doi.org/10.1002/jemt.20092

    • PubMed
    • Google Scholar
33. 1. Frankel EB
    2. Shankar R
    3. Moresco JJ
    4. Yates JR
    5. Volkmann N
    6. Audhya A

    (2017) Ist1 regulates ESCRT-III assembly and function during multivesicular endosome biogenesis in Caenorhabditis elegans embryos

    Nature Communications 8:1439.

    https://doi.org/10.1038/s41467-017-01636-8

    • PubMed
    • Google Scholar
34. 1. Freibaum BD
    2. Lu Y
    3. Lopez-Gonzalez R
    4. Kim NC
    5. Almeida S
    6. Lee KH
    7. Badders N
    8. Valentine M
    9. Miller BL
    10. Wong PC
    11. Petrucelli L
    12. Kim HJ
    13. Gao FB
    14. Taylor JP

    (2015) GGGGCC repeat expansion in C9orf72 compromises nucleocytoplasmic transport

    Nature 525:129–133.

    https://doi.org/10.1038/nature14974

    • PubMed
    • Google Scholar
35. 1. Gong YN
    2. Guy C
    3. Olauson H
    4. Becker JU
    5. Yang M
    6. Fitzgerald P
    7. Linkermann A
    8. Green DR

    (2017) ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences

    Cell 169:286–300.

    https://doi.org/10.1016/j.cell.2017.03.020

    • PubMed
    • Google Scholar
36. 1. Gonzalez Y
    2. Saito A
    3. Sazer S

    (2012) Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina

    Nucleus 3:60–76.

    https://doi.org/10.4161/nucl.18824

    • PubMed
    • Google Scholar
37. 1. Goodchild RE
    2. Kim CE
    3. Dauer WT

    (2005) Loss of the dystonia-associated protein torsinA selectively disrupts the neuronal nuclear envelope

    Neuron 48:923–932.

    https://doi.org/10.1016/j.neuron.2005.11.010

    • PubMed
    • Google Scholar
38. 1. Grund SE
    2. Fischer T
    3. Cabal GG
    4. Antúnez O
    5. Pérez-Ortín JE
    6. Hurt E

    (2008) The inner nuclear membrane protein Src1 associates with subtelomeric genes and alters their regulated gene expression

    The Journal of Cell Biology 182:897–910.

    https://doi.org/10.1083/jcb.200803098

    • PubMed
    • Google Scholar
39. 1. Gu M
    2. LaJoie D
    3. Chen OS
    4. von Appen A
    5. Ladinsky MS
    6. Redd MJ
    7. Nikolova L
    8. Bjorkman PJ
    9. Sundquist WI
    10. Ullman KS
    11. Frost A

    (2017) LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in fission yeast and human cells

    PNAS 114:E2166–E2175.

    https://doi.org/10.1073/pnas.1613916114

    • PubMed
    • Google Scholar
40. 1. Han H
    2. Monroe N
    3. Votteler J
    4. Shakya B
    5. Sundquist WI
    6. Hill CP

    (2015) Binding of substrates to the central pore of the Vps4 ATPase is autoinhibited by the microtubule interacting and trafficking (MIT) Domain and activated by MIT interacting motifs (MIMs)

    Journal of Biological Chemistry 290:13490–13499.

    https://doi.org/10.1074/jbc.M115.642355

    • PubMed
    • Google Scholar
41. 1. Han H
    2. Monroe N
    3. Sundquist WI
    4. Shen PS
    5. Hill CP

    (2017) The AAA ATPase Vps4 binds ESCRT-III substrates through a repeating array of dipeptide-binding pockets

    eLife 6:e31324.

    https://doi.org/10.7554/eLife.31324

    • PubMed
    • Google Scholar
42. 1. Hart T
    2. Chandrashekhar M
    3. Aregger M
    4. Steinhart Z
    5. Brown KR
    6. MacLeod G
    7. Mis M
    8. Zimmermann M
    9. Fradet-Turcotte A
    10. Sun S
    11. Mero P
    12. Dirks P
    13. Sidhu S
    14. Roth FP
    15. Rissland OS
    16. Durocher D
    17. Angers S
    18. Moffat J

    (2015) High-Resolution CRISPR screens reveal fitness genes and Genotype-Specific cancer liabilities

    Cell 163:1515–1526.

    https://doi.org/10.1016/j.cell.2015.11.015

    • PubMed
    • Google Scholar
43. 1. Hatch EM
    2. Fischer AH
    3. Deerinck TJ
    4. Hetzer MW

    (2013) Catastrophic nuclear envelope collapse in cancer cell micronuclei

    Cell 154:47–60.

    https://doi.org/10.1016/j.cell.2013.06.007

    • PubMed
    • Google Scholar
44. 1. Hatch E
    2. Hetzer M

    (2014) Breaching the nuclear envelope in development and disease

    The Journal of Cell Biology 205:133–141.

    https://doi.org/10.1083/jcb.201402003

    • PubMed
    • Google Scholar
45. 1. Hatch EM
    2. Hetzer MW

    (2016) Nuclear envelope rupture is induced by actin-based nucleus confinement

    The Journal of Cell Biology 215:27–36.

    https://doi.org/10.1083/jcb.201603053

    • PubMed
    • Google Scholar
46. 1. Henne WM
    2. Buchkovich NJ
    3. Zhao Y
    4. Emr SD

    (2012) The endosomal sorting complex ESCRT-II mediates the assembly and architecture of ESCRT-III helices

    Cell 151:356–371.

    https://doi.org/10.1016/j.cell.2012.08.039

    • PubMed
    • Google Scholar
47. 1. Hodge CA
    2. Choudhary V
    3. Wolyniak MJ
    4. Scarcelli JJ
    5. Schneiter R
    6. Cole CN

    (2010) Integral membrane proteins Brr6 and Apq12 link assembly of the nuclear pore complex to lipid homeostasis in the endoplasmic reticulum

    Journal of Cell Science 123:141–151.

    https://doi.org/10.1242/jcs.055046

    • PubMed
    • Google Scholar
48. 1. Horii M
    2. Shibata H
    3. Kobayashi R
    4. Katoh K
    5. Yorikawa C
    6. Yasuda J
    7. Maki M

    (2006) CHMP7, a novel ESCRT-III-related protein, associates with CHMP4b and functions in the endosomal sorting pathway

    Biochemical Journal 400:23–32.

    https://doi.org/10.1042/BJ20060897

    • PubMed
    • Google Scholar
49. 1. Jackson CE
    2. Scruggs BS
    3. Schaffer JE
    4. Hanson PI

    (2017) Effects of inhibiting VPS4 support a general role for ESCRTs in extracellular vesicle biogenesis

    Biophysical Journal 113:1342–1352.

    https://doi.org/10.1016/j.bpj.2017.05.032

    • PubMed
    • Google Scholar
50. 1. Janssens GE
    2. Meinema AC
    3. González J
    4. Wolters JC
    5. Schmidt A
    6. Guryev V
    7. Bischoff R
    8. Wit EC
    9. Veenhoff LM
    10. Heinemann M

    (2015) Protein biogenesis machinery is a driver of replicative aging in yeast

    eLife 4:e08527.

    https://doi.org/10.7554/eLife.08527

    • PubMed
    • Google Scholar
51. 1. Jevtić P
    2. Edens LJ
    3. Vuković LD
    4. Levy DL

    (2014) Sizing and shaping the nucleus: mechanisms and significance

    Current Opinion in Cell Biology 28:16–27.

    https://doi.org/10.1016/j.ceb.2014.01.003

    • PubMed
    • Google Scholar
52. 1. Jimenez AJ
    2. Maiuri P
    3. Lafaurie-Janvore J
    4. Divoux S
    5. Piel M
    6. Perez F

    (2014) ESCRT machinery is required for plasma membrane repair

    Science 343:1247136.

    https://doi.org/10.1126/science.1247136

    • PubMed
    • Google Scholar
53. 1. Jokhi V
    2. Ashley J
    3. Nunnari J
    4. Noma A
    5. Ito N
    6. Wakabayashi-Ito N
    7. Moore MJ
    8. Budnik V

    (2013) Torsin mediates primary envelopment of large ribonucleoprotein granules at the nuclear envelope

    Cell Reports 3:988–995.

    https://doi.org/10.1016/j.celrep.2013.03.015

    • PubMed
    • Google Scholar
54. 1. Jovičić A
    2. Mertens J
    3. Boeynaems S
    4. Bogaert E
    5. Chai N
    6. Yamada SB
    7. Paul JW
    8. Sun S
    9. Herdy JR
    10. Bieri G
    11. Kramer NJ
    12. Gage FH
    13. Van Den Bosch L
    14. Robberecht W
    15. Gitler AD

    (2015) Modifiers of C9orf72 dipeptide repeat toxicity connect nucleocytoplasmic transport defects to FTD/ALS

    Nature Neuroscience 18:1226–1229.

    https://doi.org/10.1038/nn.4085

    • PubMed
    • Google Scholar
55. 1. Kaneb HM
    2. Folkmann AW
    3. Belzil VV
    4. Jao LE
    5. Leblond CS
    6. Girard SL
    7. Daoud H
    8. Noreau A
    9. Rochefort D
    10. Hince P
    11. Szuto A
    12. Levert A
    13. Vidal S
    14. André-Guimont C
    15. Camu W
    16. Bouchard JP
    17. Dupré N
    18. Rouleau GA
    19. Wente SR
    20. Dion PA

    (2015) Deleterious mutations in the essential mRNA metabolism factor, hGle1, in amyotrophic lateral sclerosis

    Human Molecular Genetics 24:1363–1373.

    https://doi.org/10.1093/hmg/ddu545

    • PubMed
    • Google Scholar
56. 1. Keller A
    2. Nesvizhskii AI
    3. Kolker E
    4. Aebersold R

    (2002) Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search

    Analytical Chemistry 74:5383–5392.

    https://doi.org/10.1021/ac025747h

    • PubMed
    • Google Scholar
57. 1. Kim SJ
    2. Fernandez-Martinez J
    3. Nudelman I
    4. Shi Y
    5. Zhang W
    6. Raveh B
    7. Herricks T
    8. Slaughter BD
    9. Hogan JA
    10. Upla P
    11. Chemmama IE
    12. Pellarin R
    13. Echeverria I
    14. Shivaraju M
    15. Chaudhury AS
    16. Wang J
    17. Williams R
    18. Unruh JR
    19. Greenberg CH
    20. Jacobs EY
    21. Yu Z
    22. de la Cruz MJ
    23. Mironska R
    24. Stokes DL
    25. Aitchison JD
    26. Jarrold MF
    27. Gerton JL
    28. Ludtke SJ
    29. Akey CW
    30. Chait BT
    31. Sali A
    32. Rout MP

    (2018) Integrative structure and functional anatomy of a nuclear pore complex

    Nature 555:475–482.

    https://doi.org/10.1038/nature26003

    • PubMed
    • Google Scholar
58. 1. Kim HJ
    2. Taylor JP

    (2017) Lost in transportation: nucleocytoplasmic transport defects in ALS and other neurodegenerative diseases

    Neuron 96:285–297.

    https://doi.org/10.1016/j.neuron.2017.07.029

    • PubMed
    • Google Scholar
59. 1. King MC
    2. Lusk CP
    3. Blobel G

    (2006) Karyopherin-mediated import of integral inner nuclear membrane proteins

    Nature 442:1003–1007.

    https://doi.org/10.1038/nature05075

    • PubMed
    • Google Scholar
60. 1. Kosinski J
    2. Mosalaganti S
    3. von Appen A
    4. Teimer R
    5. DiGuilio AL
    6. Wan W
    7. Bui KH
    8. Hagen WJ
    9. Briggs JA
    10. Glavy JS
    11. Hurt E
    12. Beck M

    (2016) Molecular architecture of the inner ring scaffold of the human nuclear pore complex

    Science 352:363–365.

    https://doi.org/10.1126/science.aaf0643

    • PubMed
    • Google Scholar
61. 1. Kremer JR
    2. Mastronarde DN
    3. McIntosh JR

    (1996) Computer visualization of three-dimensional image data using IMOD

    Journal of Structural Biology 116:71–76.

    https://doi.org/10.1006/jsbi.1996.0013

    • PubMed
    • Google Scholar
62. 1. Kukulski W
    2. Schorb M
    3. Welsch S
    4. Picco A
    5. Kaksonen M
    6. Briggs JA

    (2012) Precise, correlated fluorescence microscopy and electron tomography of lowicryl sections using fluorescent fiducial markers

    Methods in Cell Biology 111:235–257.

    https://doi.org/10.1016/B978-0-12-416026-2.00013-3

    • PubMed
    • Google Scholar
63. 1. Kulak NA
    2. Pichler G
    3. Paron I
    4. Nagaraj N
    5. Mann M

    (2014) Minimal, encapsulated proteomic-sample processing applied to copy-number estimation in eukaryotic cells

    Nature Methods 11:319–324.

    https://doi.org/10.1038/nmeth.2834

    • PubMed
    • Google Scholar
64. 1. LaCava J
    2. Fernandez-Martinez J
    3. Hakhverdyan Z
    4. Rout MP

    (2016) Optimized affinity capture of yeast protein complexes

    Cold Spring Harbor Protocols 2016:prot087932.

    https://doi.org/10.1101/pdb.prot087932

    • Google Scholar
65. 1. Lata S
    2. Roessle M
    3. Solomons J
    4. Jamin M
    5. Gottlinger HG
    6. Svergun DI
    7. Weissenhorn W

    (2008) Structural basis for autoinhibition of ESCRT-III CHMP3

    Journal of Molecular Biology 378:818–827.

    https://doi.org/10.1016/j.jmb.2008.03.030

    • PubMed
    • Google Scholar
66. 1. Laudermilch E
    2. Tsai PL
    3. Graham M
    4. Turner E
    5. Zhao C
    6. Schlieker C

    (2016) Dissecting torsin/cofactor function at the nuclear envelope: a genetic study

    Molecular Biology of the Cell 27:3964–3971.

    https://doi.org/10.1091/mbc.e16-07-0511

    • PubMed
    • Google Scholar
67. 1. Lee CP
    2. Liu PT
    3. Kung HN
    4. Su MT
    5. Chua HH
    6. Chang YH
    7. Chang CW
    8. Tsai CH
    9. Liu FT
    10. Chen MR

    (2012) The ESCRT machinery is recruited by the viral BFRF1 protein to the nucleus-associated membrane for the maturation of Epstein-Barr virus

    PLOS Pathogens 8:e1002904.

    https://doi.org/10.1371/journal.ppat.1002904

    • PubMed
    • Google Scholar
68. 1. Lee CP
    2. Liu GT
    3. Kung HN
    4. Liu PT
    5. Liao YT
    6. Chow LP
    7. Chang LS
    8. Chang YH
    9. Chang CW
    10. Shu WC
    11. Angers A
    12. Farina A
    13. Lin SF
    14. Tsai CH
    15. Bouamr F
    16. Chen MR

    (2016) The ubiquitin ligase itch and ubiquitination regulate BFRF1-Mediated nuclear envelope modification for Epstein-Barr virus maturation

    Journal of Virology 90:8994–9007.

    https://doi.org/10.1128/JVI.01235-16

    • PubMed
    • Google Scholar
69. 1. Li X
    2. Qian J
    3. Wang C
    4. Zheng K
    5. Ye L
    6. Fu Y
    7. Han N
    8. Bian H
    9. Pan J
    10. Wang J
    11. Zhu M

    (2011) Regulating cytoplasmic calcium homeostasis can reduce aluminum toxicity in yeast

    PLOS ONE 6:e21148.

    https://doi.org/10.1371/journal.pone.0021148

    • PubMed
    • Google Scholar
70. 1. Lokareddy RK
    2. Hapsari RA
    3. van Rheenen M
    4. Pumroy RA
    5. Bhardwaj A
    6. Steen A
    7. Veenhoff LM
    8. Cingolani G

    (2015) Distinctive properties of the nuclear localization signals of inner nuclear membrane proteins Heh1 and Heh2

    Structure 23:1305–1316.

    https://doi.org/10.1016/j.str.2015.04.017

    • PubMed
    • Google Scholar
71. 1. Longtine MS
    2. McKenzie A
    3. Demarini DJ
    4. Shah NG
    5. Wach A
    6. Brachat A
    7. Philippsen P
    8. Pringle JR

    (1998) Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae

    Yeast 14:953–961.

    https://doi.org/10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U

    • PubMed
    • Google Scholar
72. 1. Lord CL
    2. Timney BL
    3. Rout MP
    4. Wente SR

    (2015) Altering nuclear pore complex function impacts longevity and mitochondrial function in S. cerevisiae

    The Journal of Cell Biology 208:729–744.

    https://doi.org/10.1083/jcb.201412024

    • PubMed
    • Google Scholar
73. 1. Lusk CP
    2. Blobel G
    3. King MC

    (2007) Highway to the inner nuclear membrane: rules for the road

    Nature Reviews Molecular Cell Biology 8:414–420.

    https://doi.org/10.1038/nrm2165

    • PubMed
    • Google Scholar
74. 1. Lusk CP
    2. King MC

    (2017) The nucleus: keeping it together by keeping it apart

    Current Opinion in Cell Biology 44:44–50.

    https://doi.org/10.1016/j.ceb.2017.02.001

    • PubMed
    • Google Scholar
75. 1. Maciejowski J
    2. Li Y
    3. Bosco N
    4. Campbell PJ
    5. de Lange T

    (2015) Chromothripsis and Kataegis induced by telomere crisis

    Cell 163:1641–1654.

    https://doi.org/10.1016/j.cell.2015.11.054

    • PubMed
    • Google Scholar
76. 1. Madrid AS
    2. Mancuso J
    3. Cande WZ
    4. Weis K

    (2006) The role of the integral membrane nucleoporins Ndc1p and Pom152p in nuclear pore complex assembly and function

    The Journal of Cell Biology 173:361–371.

    https://doi.org/10.1083/jcb.200506199

    • PubMed
    • Google Scholar
77. 1. Makio T
    2. Lapetina DL
    3. Wozniak RW

    (2013) Inheritance of yeast nuclear pore complexes requires the Nsp1p subcomplex

    The Journal of Cell Biology 203:187–196.

    https://doi.org/10.1083/jcb.201304047

    • PubMed
    • Google Scholar
78. 1. Malhas A
    2. Goulbourne C
    3. Vaux DJ

    (2011) The nucleoplasmic reticulum: form and function

    Trends in Cell Biology 21:362–373.

    https://doi.org/10.1016/j.tcb.2011.03.008

    • PubMed
    • Google Scholar
79. 1. Mans B
    2. Anantharaman V
    3. Aravind L
    4. Koonin EV

    (2004) Comparative genomics, evolution and origins of the nuclear envelope and nuclear pore complex

    Cell Cycle 3:1625–1650.

    https://doi.org/10.4161/cc.3.12.1316

    • Google Scholar
80. 1. Mastronarde DN

    (2005) Automated electron microscope tomography using robust prediction of specimen movements

    Journal of Structural Biology 152:36–51.

    https://doi.org/10.1016/j.jsb.2005.07.007

    • PubMed
    • Google Scholar
81. 1. Maul GG
    2. Maul HM
    3. Scogna JE
    4. Lieberman MW
    5. Stein GS
    6. Hsu BY
    7. Borun TW

    (1972) Time sequence of nuclear pore formation in phytohemagglutinin-stimulated lymphocytes and in HeLa cells during the cell cycle

    The Journal of Cell Biology 55:433–447.

    https://doi.org/10.1083/jcb.55.2.433

    • PubMed
    • Google Scholar
82. 1. McCullough J
    2. Clippinger AK
    3. Talledge N
    4. Skowyra ML
    5. Saunders MG
    6. Naismith TV
    7. Colf LA
    8. Afonine P
    9. Arthur C
    10. Sundquist WI
    11. Hanson PI
    12. Frost A

    (2015) Structure and membrane remodeling activity of ESCRT-III helical polymers

    Science 350:1548–1551.

    https://doi.org/10.1126/science.aad8305

    • PubMed
    • Google Scholar
83. 1. McCullough J
    2. Frost A
    3. Sundquist WI

    (2018) Structures, functions, and dynamics of ESCRT-III/Vps4 membrane remodeling and fission complexes

    Annual Review of Cell and Developmental Biology 34:85–109.

    https://doi.org/10.1146/annurev-cellbio-100616-060600

    • PubMed
    • Google Scholar
84. 1. Meinema AC
    2. Laba JK
    3. Hapsari RA
    4. Otten R
    5. Mulder FA
    6. Kralt A
    7. van den Bogaart G
    8. Lusk CP
    9. Poolman B
    10. Veenhoff LM

    (2011) Long unfolded linkers facilitate membrane protein import through the nuclear pore complex

    Science 333:90–93.

    https://doi.org/10.1126/science.1205741

    • PubMed
    • Google Scholar
85. 1. Mekhail K
    2. Seebacher J
    3. Gygi SP
    4. Moazed D

    (2008) Role for perinuclear chromosome tethering in maintenance of genome stability

    Nature 456:667–670.

    https://doi.org/10.1038/nature07460

    • PubMed
    • Google Scholar
86. 1. Miyake N
    2. Tsukaguchi H
    3. Koshimizu E
    4. Shono A
    5. Matsunaga S
    6. Shiina M
    7. Mimura Y
    8. Imamura S
    9. Hirose T
    10. Okudela K
    11. Nozu K
    12. Akioka Y
    13. Hattori M
    14. Yoshikawa N
    15. Kitamura A
    16. Cheong HI
    17. Kagami S
    18. Yamashita M
    19. Fujita A
    20. Miyatake S
    21. Tsurusaki Y
    22. Nakashima M
    23. Saitsu H
    24. Ohashi K
    25. Imamoto N
    26. Ryo A
    27. Ogata K
    28. Iijima K
    29. Matsumoto N

    (2015) Biallelic mutations in nuclear pore complex subunit NUP107 cause Early-Childhood-Onset Steroid-Resistant nephrotic syndrome

    The American Journal of Human Genetics 97:555–566.

    https://doi.org/10.1016/j.ajhg.2015.08.013

    • PubMed
    • Google Scholar
87. 1. Mochida K
    2. Oikawa Y
    3. Kimura Y
    4. Kirisako H
    5. Hirano H
    6. Ohsumi Y
    7. Nakatogawa H

    (2015) Receptor-mediated selective autophagy degrades the endoplasmic reticulum and the nucleus

    Nature 522:359–362.

    https://doi.org/10.1038/nature14506

    • PubMed
    • Google Scholar
88. 1. Monroe N
    2. Han H
    3. Shen PS
    4. Sundquist WI
    5. Hill CP

    (2017) Structural basis of protein translocation by the Vps4-Vta1 AAA ATPase

    eLife 6:e24487.

    https://doi.org/10.7554/eLife.24487

    • PubMed
    • Google Scholar
89. 1. Morita E
    2. Sandrin V
    3. McCullough J
    4. Katsuyama A
    5. Baci Hamilton I
    6. Sundquist WI

    (2011) ESCRT-III protein requirements for HIV-1 budding

    Cell Host & Microbe 9:235–242.

    https://doi.org/10.1016/j.chom.2011.02.004

    • PubMed
    • Google Scholar
90. 1. Mostofa MG
    2. Rahman MA
    3. Koike N
    4. Yeasmin AM
    5. Islam N
    6. Waliullah TM
    7. Hosoyamada S
    8. Shimobayashi M
    9. Kobayashi T
    10. Hall MN
    11. Ushimaru T

    (2018) CLIP and cohibin separate rDNA from nucleolar proteins destined for degradation by nucleophagy

    The Journal of Cell Biology 217:2675–2690.

    https://doi.org/10.1083/jcb.201706164

    • PubMed
    • Google Scholar
91. 1. Nesvizhskii AI
    2. Keller A
    3. Kolker E
    4. Aebersold R

    (2003) A statistical model for identifying proteins by tandem mass spectrometry

    Analytical Chemistry 75:4646–4658.

    https://doi.org/10.1021/ac0341261

    • PubMed
    • Google Scholar
92. 1. Neville M
    2. Rosbash M

    (1999) The NES-Crm1p export pathway is not a major mRNA export route in Saccharomyces cerevisiae

    The EMBO Journal 18:3746–3756.

    https://doi.org/10.1093/emboj/18.13.3746

    • PubMed
    • Google Scholar
93. 1. Nousiainen HO
    2. Kestilä M
    3. Pakkasjärvi N
    4. Honkala H
    5. Kuure S
    6. Tallila J
    7. Vuopala K
    8. Ignatius J
    9. Herva R
    10. Peltonen L

    (2008) Mutations in mRNA export mediator GLE1 result in a fetal motoneuron disease

    Nature Genetics 40:155–157.

    https://doi.org/10.1038/ng.2007.65

    • PubMed
    • Google Scholar
94. 1. Obita T
    2. Saksena S
    3. Ghazi-Tabatabai S
    4. Gill DJ
    5. Perisic O
    6. Emr SD
    7. Williams RL

    (2007) Structural basis for selective recognition of ESCRT-III by the AAA ATPase Vps4

    Nature 449:735–739.

    https://doi.org/10.1038/nature06171

    • PubMed
    • Google Scholar
95. 1. Oeffinger M
    2. Wei KE
    3. Rogers R
    4. DeGrasse JA
    5. Chait BT
    6. Aitchison JD
    7. Rout MP

    (2007) Comprehensive analysis of diverse ribonucleoprotein complexes

    Nature Methods 4:951–956.

    https://doi.org/10.1038/nmeth1101

    • PubMed
    • Google Scholar
96. 1. Olmos Y
    2. Hodgson L
    3. Mantell J
    4. Verkade P
    5. Carlton JG

    (2015) ESCRT-III controls nuclear envelope reformation

    Nature 522:236–239.

    https://doi.org/10.1038/nature14503

    • PubMed
    • Google Scholar
97. 1. Olmos Y
    2. Perdrix-Rosell A
    3. Carlton JG

    (2016) Membrane binding by CHMP7 coordinates ESCRT-III-Dependent nuclear envelope reformation

    Current Biology 26:2635–2641.

    https://doi.org/10.1016/j.cub.2016.07.039

    • PubMed
    • Google Scholar
98. 1. Onischenko E
    2. Tang JH
    3. Andersen KR
    4. Knockenhauer KE
    5. Vallotton P
    6. Derrer CP
    7. Kralt A
    8. Mugler CF
    9. Chan LY
    10. Schwartz TU
    11. Weis K

    (2017) Natively unfolded FG repeats stabilize the structure of the nuclear pore complex

    Cell 171:904–917.

    https://doi.org/10.1016/j.cell.2017.09.033

    • PubMed
    • Google Scholar
99. 1. Otsuka S
    2. Bui KH
    3. Schorb M
    4. Hossain MJ
    5. Politi AZ
    6. Koch B
    7. Eltsov M
    8. Beck M
    9. Ellenberg J

    (2016) Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope

    eLife 5:e19017.

    https://doi.org/10.7554/eLife.19071

    • Google Scholar
100. 1. Pappas SS
     2. Liang CC
     3. Kim S
     4. Rivera CO
     5. Dauer WT

     (2018) TorsinA dysfunction causes persistent neuronal nuclear pore defects

     Human Molecular Genetics 27:407–420.

     https://doi.org/10.1093/hmg/ddx405

     • PubMed
     • Google Scholar
101. 1. Paul-Gilloteaux P
     2. Heiligenstein X
     3. Belle M
     4. Domart MC
     5. Larijani B
     6. Collinson L
     7. Raposo G
     8. Salamero J

     (2017) eC-CLEM: flexible multidimensional registration software for correlative microscopies

     Nature Methods 14:102–103.

     https://doi.org/10.1038/nmeth.4170

     • PubMed
     • Google Scholar
102. 1. Popken P
     2. Ghavami A
     3. Onck PR
     4. Poolman B
     5. Veenhoff LM

     (2015) Size-dependent leak of soluble and membrane proteins through the yeast nuclear pore complex

     Molecular Biology of the Cell 26:1386–1394.

     https://doi.org/10.1091/mbc.E14-07-1175

     • PubMed
     • Google Scholar
103. 1. Raab M
     2. Gentili M
     3. de Belly H
     4. Thiam HR
     5. Vargas P
     6. Jimenez AJ
     7. Lautenschlaeger F
     8. Voituriez R
     9. Lennon-Duménil AM
     10. Manel N
     11. Piel M

     (2016) ESCRT III repairs nuclear envelope ruptures during cell migration to limit DNA damage and cell death

     Science 352:359–362.

     https://doi.org/10.1126/science.aad7611

     • PubMed
     • Google Scholar
104. 1. Radulovic M
     2. Schink KO
     3. Wenzel EM
     4. Nähse V
     5. Bongiovanni A
     6. Lafont F
     7. Stenmark H

     (2018) ESCRT-mediated lysosome repair precedes lysophagy and promotes cell survival

     The EMBO Journal 37:e99753.

     https://doi.org/10.15252/embj.201899753

     • PubMed
     • Google Scholar
105. 1. Roberts P
     2. Moshitch-Moshkovitz S
     3. Kvam E
     4. O'Toole E
     5. Winey M
     6. Goldfarb DS

     (2003) Piecemeal microautophagy of nucleus in Saccharomyces cerevisiae

     Molecular Biology of the Cell 14:129–141.

     https://doi.org/10.1091/mbc.e02-08-0483

     • PubMed
     • Google Scholar
106. 1. Robijns J
     2. Molenberghs F
     3. Sieprath T
     4. Corne TD
     5. Verschuuren M
     6. De Vos WH

     (2016) In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

     Scientific Reports 6:30325.

     https://doi.org/10.1038/srep30325

     • PubMed
     • Google Scholar
107. 1. Romanauska A
     2. Köhler A

     (2018) The inner nuclear membrane is a metabolically active territory that generates nuclear lipid droplets

     Cell 174:700–715.

     https://doi.org/10.1016/j.cell.2018.05.047

     • PubMed
     • Google Scholar
108. 1. Rue SM
     2. Mattei S
     3. Saksena S
     4. Emr SD

     (2008) Novel Ist1-Did2 complex functions at a late step in Multivesicular body sorting

     Molecular Biology of the Cell 19:475–484.

     https://doi.org/10.1091/mbc.e07-07-0694

     • PubMed
     • Google Scholar
109. 1. Saksena S
     2. Wahlman J
     3. Teis D
     4. Johnson AE
     5. Emr SD

     (2009) Functional reconstitution of ESCRT-III assembly and disassembly

     Cell 136:97–109.

     https://doi.org/10.1016/j.cell.2008.11.013

     • PubMed
     • Google Scholar
110. 1. Savas JN
     2. Toyama BH
     3. Xu T
     4. Yates JR
     5. Hetzer MW

     (2012) Extremely long-lived nuclear pore proteins in the rat brain

     Science 335:942.

     https://doi.org/10.1126/science.1217421

     • PubMed
     • Google Scholar
111. 1. Scarcelli JJ
     2. Hodge CA
     3. Cole CN

     (2007) The yeast integral membrane protein Apq12 potentially links membrane dynamics to assembly of nuclear pore complexes

     The Journal of Cell Biology 178:799–812.

     https://doi.org/10.1083/jcb.200702120

     • PubMed
     • Google Scholar
112. 1. Scheffer LL
     2. Sreetama SC
     3. Sharma N
     4. Medikayala S
     5. Brown KJ
     6. Defour A
     7. Jaiswal JK

     (2014) Mechanism of Ca²⁺-triggered ESCRT assembly and regulation of cell membrane repair

     Nature Communications 5:5646.

     https://doi.org/10.1038/ncomms6646

     • PubMed
     • Google Scholar
113. 1. Schindelin J
     2. Arganda-Carreras I
     3. Frise E
     4. Kaynig V
     5. Longair M
     6. Pietzsch T
     7. Preibisch S
     8. Rueden C
     9. Saalfeld S
     10. Schmid B
     11. Tinevez JY
     12. White DJ
     13. Hartenstein V
     14. Eliceiri K
     15. Tomancak P
     16. Cardona A

     (2012) Fiji: an open-source platform for biological-image analysis

     Nature Methods 9:676–682.

     https://doi.org/10.1038/nmeth.2019

     • PubMed
     • Google Scholar
114. 1. Schmidt HB
     2. Görlich D

     (2016) Transport selectivity of nuclear pores, phase separation, and membraneless organelles

     Trends in Biochemical Sciences 41:46–61.

     https://doi.org/10.1016/j.tibs.2015.11.001

     • PubMed
     • Google Scholar
115. 1. Schneiter R
     2. Hitomi M
     3. Ivessa AS
     4. Fasch EV
     5. Kohlwein SD
     6. Tartakoff AM

     (1996) A yeast acetyl coenzyme A carboxylase mutant links very-long-chain fatty acid synthesis to the structure and function of the nuclear membrane-pore complex

     Molecular and Cellular Biology 16:7161–7172.

     https://doi.org/10.1128/MCB.16.12.7161

     • PubMed
     • Google Scholar
116. 1. Schöneberg J
     2. Lee IH
     3. Iwasa JH
     4. Hurley JH

     (2017) Reverse-topology membrane scission by the ESCRT proteins

     Nature Reviews Molecular Cell Biology 18:5–17.

     https://doi.org/10.1038/nrm.2016.121

     • PubMed
     • Google Scholar
117. 1. Schöneberg J
     2. Pavlin MR
     3. Yan S
     4. Righini M
     5. Lee IH
     6. Carlson LA
     7. Bahrami AH
     8. Goldman DH
     9. Ren X
     10. Hummer G
     11. Bustamante C
     12. Hurley JH

     (2018) ATP-dependent force generation and membrane scission by ESCRT-III and Vps4

     Science 362:1423–1428.

     https://doi.org/10.1126/science.aat1839

     • PubMed
     • Google Scholar
118. 1. Schreiber KH
     2. Kennedy BK

     (2013) When lamins go bad: nuclear structure and disease

     Cell 152:1365–1375.

     https://doi.org/10.1016/j.cell.2013.02.015

     • PubMed
     • Google Scholar
119. 1. Schreiner SM
     2. Koo PK
     3. Zhao Y
     4. Mochrie SG
     5. King MC

     (2015) The tethering of chromatin to the nuclear envelope supports nuclear mechanics

     Nature Communications 6:7159.

     https://doi.org/10.1038/ncomms8159

     • PubMed
     • Google Scholar
120. 1. Searle BC
     2. Turner M
     3. Nesvizhskii AI

     (2008) Improving sensitivity by probabilistically combining results from multiple MS/MS search methodologies

     Journal of Proteome Research 7:245–253.

     https://doi.org/10.1021/pr070540w

     • PubMed
     • Google Scholar
121. 1. Serebryannyy L
     2. Misteli T

     (2018) Protein sequestration at the nuclear periphery as a potential regulatory mechanism in premature aging

     The Journal of Cell Biology 217:21–37.

     https://doi.org/10.1083/jcb.201706061

     • PubMed
     • Google Scholar
122. 1. Shestakova A
     2. Hanono A
     3. Drosner S
     4. Curtiss M
     5. Davies BA
     6. Katzmann DJ
     7. Babst M

     (2010) Assembly of the AAA ATPase Vps4 on ESCRT-III

     Molecular Biology of the Cell 21:1059–1071.

     https://doi.org/10.1091/mbc.e09-07-0572

     • PubMed
     • Google Scholar
123. 1. Shi KY
     2. Mori E
     3. Nizami ZF
     4. Lin Y
     5. Kato M
     6. Xiang S
     7. Wu LC
     8. Ding M
     9. Yu Y
     10. Gall JG
     11. McKnight SL

     (2017) Toxic PR_n poly-dipeptides encoded by the C9orf72 repeat expansio_n block nuclear import and export

     PNAS 114:E1111–E1117.

     https://doi.org/10.1073/pnas.1620293114

     • PubMed
     • Google Scholar
124. 1. Shim S
     2. Kimpler LA
     3. Hanson PI

     (2007) Structure/function analysis of four core ESCRT-III proteins reveals common regulatory role for extreme C-terminal domain

     Traffic 8:1068–1079.

     https://doi.org/10.1111/j.1600-0854.2007.00584.x

     • PubMed
     • Google Scholar
125. 1. Skowyra ML
     2. Schlesinger PH
     3. Naismith TV
     4. Hanson PI

     (2018) Triggered recruitment of ESCRT machinery promotes endolysosomal repair

     Science 360:eaar5078.

     https://doi.org/10.1126/science.aar5078

     • PubMed
     • Google Scholar
126. 1. Speese SD
     2. Ashley J
     3. Jokhi V
     4. Nunnari J
     5. Barria R
     6. Li Y
     7. Ataman B
     8. Koon A
     9. Chang YT
     10. Li Q
     11. Moore MJ
     12. Budnik V

     (2012) Nuclear envelope budding enables large ribonucleoprotein particle export during synaptic wnt signaling

     Cell 149:832–846.

     https://doi.org/10.1016/j.cell.2012.03.032

     • PubMed
     • Google Scholar
127. 1. Stuchell-Brereton MD
     2. Skalicky JJ
     3. Kieffer C
     4. Karren MA
     5. Ghaffarian S
     6. Sundquist WI

     (2007) ESCRT-III recognition by VPS4 ATPases

     Nature 449:740–744.

     https://doi.org/10.1038/nature06172

     • PubMed
     • Google Scholar
128. 1. Su M
     2. Guo EZ
     3. Ding X
     4. Li Y
     5. Tarrasch JT
     6. Brooks CL
     7. Xu Z
     8. Skiniotis G

     (2017) Mechanism of Vps4 hexamer function revealed by cryo-EM

     Science Advances 3:e1700325.

     https://doi.org/10.1126/sciadv.1700325

     • PubMed
     • Google Scholar
129. 1. Tang S
     2. Henne WM
     3. Borbat PP
     4. Buchkovich NJ
     5. Freed JH
     6. Mao Y
     7. Fromme JC
     8. Emr SD

     (2015) Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments

     eLife 4:e12548.

     https://doi.org/10.7554/eLife.12548

     • PubMed
     • Google Scholar
130. 1. Tang S
     2. Buchkovich NJ
     3. Henne WM
     4. Banjade S
     5. Kim YJ
     6. Emr SD

     (2016) ESCRT-III activation by parallel action of ESCRT-I/II and ESCRT-0/Bro1 during MVB biogenesis

     eLife 5:e15507.

     https://doi.org/10.7554/eLife.15507

     • PubMed
     • Google Scholar
131. 1. Teis D
     2. Saksena S
     3. Emr SD

     (2008) Ordered assembly of the ESCRT-III complex on endosomes is required to sequester cargo during MVB formation

     Developmental Cell 15:578–589.

     https://doi.org/10.1016/j.devcel.2008.08.013

     • PubMed
     • Google Scholar
132. 1. Teis D
     2. Saksena S
     3. Judson BL
     4. Emr SD

     (2010) ESCRT-II coordinates the assembly of ESCRT-III filaments for cargo sorting and multivesicular body vesicle formation

     The EMBO Journal 29:871–883.

     https://doi.org/10.1038/emboj.2009.408

     • PubMed
     • Google Scholar
133. 1. Thaller DJ
     2. Lusk CP

     (2018) Fantastic nuclear envelope herniations and where to find them

     Biochemical Society Transactions 46:877–889.

     https://doi.org/10.1042/BST20170442

     • Google Scholar
134. 1. Timney BL
     2. Raveh B
     3. Mironska R
     4. Trivedi JM
     5. Kim SJ
     6. Russel D
     7. Wente SR
     8. Sali A
     9. Rout MP

     (2016) Simple rules for passive diffusion through the nuclear pore complex

     The Journal of Cell Biology 215:57–76.

     https://doi.org/10.1083/jcb.201601004

     • PubMed
     • Google Scholar
135. 1. Toyama BH
     2. Savas JN
     3. Park SK
     4. Harris MS
     5. Ingolia NT
     6. Yates JR
     7. Hetzer MW

     (2013) Identification of Long-Lived proteins reveals exceptional stability of essential cellular structures

     Cell 154:971–982.

     https://doi.org/10.1016/j.cell.2013.07.037

     • Google Scholar
136. 1. Toyama BH
     2. Arrojo E Drigo R
     3. Lev-Ram V
     4. Ramachandra R
     5. Deerinck TJ
     6. Lechene C
     7. Ellisman MH
     8. Hetzer MW

     (2019) Visualization of long-lived proteins reveals age mosaicism within nuclei of postmitotic cells

     The Journal of Cell Biology 218:433–444.

     https://doi.org/10.1083/jcb.201809123

     • PubMed
     • Google Scholar
137. 1. Van Driessche B
     2. Tafforeau L
     3. Hentges P
     4. Carr AM
     5. Vandenhaute J

     (2005) Additional vectors for PCR-based gene tagging in Saccharomyces cerevisiae and Schizosaccharomyces pombe using nourseothricin resistance

     Yeast 22:1061–1068.

     https://doi.org/10.1002/yea.1293

     • PubMed
     • Google Scholar
138. 1. Vargas JD
     2. Hatch EM
     3. Anderson DJ
     4. Hetzer MW

     (2012) Transient nuclear envelope rupturing during interphase in human cancer cells

     Nucleus 3:88–100.

     https://doi.org/10.4161/nucl.18954

     • PubMed
     • Google Scholar
139. 1. Ventimiglia LN
     2. Cuesta-Geijo MA
     3. Martinelli N
     4. Caballe A
     5. Macheboeuf P
     6. Miguet N
     7. Parnham IM
     8. Olmos Y
     9. Carlton JG
     10. Weissenhorn W
     11. Martin-Serrano J

     (2018) CC2D1B coordinates ESCRT-III activity during the mitotic reformation of the nuclear envelope

     Developmental Cell 47:547–563.

     https://doi.org/10.1016/j.devcel.2018.11.012

     • PubMed
     • Google Scholar
140. 1. Vietri M
     2. Schink KO
     3. Campsteijn C
     4. Wegner CS
     5. Schultz SW
     6. Christ L
     7. Thoresen SB
     8. Brech A
     9. Raiborg C
     10. Stenmark H

     (2015) Spastin and ESCRT-III coordinate mitotic spindle disassembly and nuclear envelope sealing

     Nature 522:231–235.

     https://doi.org/10.1038/nature14408

     • PubMed
     • Google Scholar
141. 1. von Schwedler UK
     2. Stuchell M
     3. Müller B
     4. Ward DM
     5. Chung HY
     6. Morita E
     7. Wang HE
     8. Davis T
     9. He GP
     10. Cimbora DM
     11. Scott A
     12. Kräusslich HG
     13. Kaplan J
     14. Morham SG
     15. Sundquist WI

     (2003) The protein network of HIV budding

     Cell 114:701–713.

     https://doi.org/10.1016/S0092-8674(03)00714-1

     • PubMed
     • Google Scholar
142. 1. Webster BM
     2. Colombi P
     3. Jäger J
     4. Lusk CP

     (2014) Surveillance of nuclear pore complex assembly by ESCRT-III/Vps4

     Cell 159:388–401.

     https://doi.org/10.1016/j.cell.2014.09.012

     • PubMed
     • Google Scholar
143. 1. Webster BM
     2. Thaller DJ
     3. Jäger J
     4. Ochmann SE
     5. Borah S
     6. Lusk CP

     (2016) Chm7 and Heh1 collaborate to link nuclear pore complex quality control with nuclear envelope sealing

     The EMBO Journal 35:2447–2467.

     https://doi.org/10.15252/embj.201694574

     • PubMed
     • Google Scholar
144. 1. Wemmer M
     2. Azmi I
     3. West M
     4. Davies B
     5. Katzmann D
     6. Odorizzi G

     (2011) Bro1 binding to Snf7 regulates ESCRT-III membrane scission activity in yeast

     The Journal of Cell Biology 192:295–306.

     https://doi.org/10.1083/jcb.201007018

     • PubMed
     • Google Scholar
145. 1. Wente SR
     2. Blobel G

     (1993) A temperature-sensitive NUP116 null mutant forms a nuclear envelope seal over the yeast nuclear pore complex thereby blocking nucleocytoplasmic traffic

     The Journal of Cell Biology 123:275–284.

     https://doi.org/10.1083/jcb.123.2.275

     • PubMed
     • Google Scholar
146. 1. Wenzel EM
     2. Schultz SW
     3. Schink KO
     4. Pedersen NM
     5. Nähse V
     6. Carlson A
     7. Brech A
     8. Stenmark H
     9. Raiborg C

     (2018) Concerted ESCRT and clathrin recruitment waves define the timing and morphology of intraluminal vesicle formation

     Nature Communications 9:2932.

     https://doi.org/10.1038/s41467-018-05345-8

     • PubMed
     • Google Scholar
147. 1. West M
     2. Zurek N
     3. Hoenger A
     4. Voeltz GK

     (2011) A 3D analysis of yeast ER structure reveals how ER domains are organized by membrane curvature

     The Journal of Cell Biology 193:333–346.

     https://doi.org/10.1083/jcb.201011039

     • PubMed
     • Google Scholar
148. 1. Winey M
     2. Yarar D
     3. Giddings TH
     4. Mastronarde DN

     (1997) Nuclear pore complex number and distribution throughout the Saccharomyces cerevisiae cell cycle by three-dimensional reconstruction from electron micrographs of nuclear envelopes

     Molecular Biology of the Cell 8:2119–2132.

     https://doi.org/10.1091/mbc.8.11.2119

     • PubMed
     • Google Scholar
149. 1. Xiao J
     2. Chen XW
     3. Davies BA
     4. Saltiel AR
     5. Katzmann DJ
     6. Xu Z

     (2009) Structural basis of Ist1 function and Ist1-Did2 interaction in the multivesicular body pathway and cytokinesis

     Molecular Biology of the Cell 20:3514–3524.

     https://doi.org/10.1091/mbc.e09-05-0403

     • PubMed
     • Google Scholar
150. 1. Xu D
     2. Marquis K
     3. Pei J
     4. Fu SC
     5. Cağatay T
     6. Grishin NV
     7. Chook YM

     (2015) LocNES: a computational tool for locating classical NESs in CRM1 cargo proteins

     Bioinformatics 31:1357–1365.

     https://doi.org/10.1093/bioinformatics/btu826

     • PubMed
     • Google Scholar
151. 1. Yam C
     2. Gu Y
     3. Oliferenko S

     (2013) Partitioning and remodeling of the schizosaccharomyces japonicus mitotic nucleus require chromosome tethers

     Current Biology 23:2303–2310.

     https://doi.org/10.1016/j.cub.2013.09.057

     • PubMed
     • Google Scholar
152. 1. Yang Z
     2. Vild C
     3. Ju J
     4. Zhang X
     5. Liu J
     6. Shen J
     7. Zhao B
     8. Lan W
     9. Gong F
     10. Liu M
     11. Cao C
     12. Xu Z

     (2012) Structural basis of molecular recognition between ESCRT-III-like protein Vps60 and AAA-ATPase regulator Vta1 in the multivesicular body pathway

     Journal of Biological Chemistry 287:43899–43908.

     https://doi.org/10.1074/jbc.M112.390724

     • PubMed
     • Google Scholar
153. 1. Yang B
     2. Stjepanovic G
     3. Shen Q
     4. Martin A
     5. Hurley JH

     (2015) Vps4 disassembles an ESCRT-III filament by global unfolding and processive translocation

     Nature Structural & Molecular Biology 22:492–498.

     https://doi.org/10.1038/nsmb.3015

     • PubMed
     • Google Scholar
154. 1. Yewdell WT
     2. Colombi P
     3. Makhnevych T
     4. Lusk CP

     (2011) Lumenal interactions in nuclear pore complex assembly and stability

     Molecular Biology of the Cell 22:1375–1388.

     https://doi.org/10.1091/mbc.e10-06-0554

     • PubMed
     • Google Scholar
155. 1. Zamborlini A
     2. Usami Y
     3. Radoshitzky SR
     4. Popova E
     5. Palu G
     6. Göttlinger H

     (2006) Release of autoinhibition converts ESCRT-III components into potent inhibitors of HIV-1 budding

     PNAS 103:19140–19145.

     https://doi.org/10.1073/pnas.0603788103

     • PubMed
     • Google Scholar
156. 1. Zhang K
     2. Donnelly CJ
     3. Haeusler AR
     4. Grima JC
     5. Machamer JB
     6. Steinwald P
     7. Daley EL
     8. Miller SJ
     9. Cunningham KM
     10. Vidensky S
     11. Gupta S
     12. Thomas MA
     13. Hong I
     14. Chiu SL
     15. Huganir RL
     16. Ostrow LW
     17. Matunis MJ
     18. Wang J
     19. Sattler R
     20. Lloyd TE
     21. Rothstein JD

     (2015) The C9orf72 repeat expansion disrupts nucleocytoplasmic transport

     Nature 525:56–61.

     https://doi.org/10.1038/nature14973

     • PubMed
     • Google Scholar
157. 1. Zhang W
     2. Neuner A
     3. Rüthnick D
     4. Sachsenheimer T
     5. Lüchtenborg C
     6. Brügger B
     7. Schiebel E

     (2018) Brr6 and Brl1 locate to nuclear pore complex assembly sites to promote their biogenesis

     The Journal of Cell Biology 217:877–894.

     https://doi.org/10.1083/jcb.201706024

     • PubMed
     • Google Scholar

Author details

1. David J Thaller

   Department of Cell Biology, Yale School of Medicine, New Haven, United States

   Contribution

   Conceptualization, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing—original draft, Writing—review and editing

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0003-3577-5562

2. Matteo Allegretti

   Structural and Computational Biology Unit, European Molecular Biology Laboratory, Meyerhofstrasse, Germany

   Contribution

   Formal analysis, Validation, Investigation, Visualization, Writing—review and editing

   Competing interests

   No competing interests declared

3. Sapan Borah

   Department of Cell Biology, Yale School of Medicine, New Haven, United States

   Contribution

   Investigation, Visualization, Writing—review and editing

   Competing interests

   No competing interests declared

4. Paolo Ronchi

   Electron Microscopy Core Facility, European Molecular Biology Laboratory, Meyerhofstrasse, Germany

   Contribution

   Resources, Investigation, Writing—review and editing

   Competing interests

   No competing interests declared

5. Martin Beck

   Structural and Computational Biology Unit, European Molecular Biology Laboratory, Meyerhofstrasse, Germany

   Contribution

   Resources, Supervision, Project administration, Writing—review and editing

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-7397-1321

6. C Patrick Lusk

   Department of Cell Biology, Yale School of Medicine, New Haven, United States

   Contribution

   Conceptualization, Resources, Formal analysis, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing—original draft, Project administration, Writing—review and editing

   For correspondence

   patrick.lusk@yale.edu

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0003-4703-0533

• 4,587

  views

• 774

  downloads

• 97

  citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.