Natural rubber (NR) is a non-fungible natural polymer used for manufacturing rubber products such as tires because of its unique physical properties. No other synthetic polymer has been developed to date, with physical properties comparable to NR. Although more than 2500 plant species are known to biosynthesize NR (Metcalfe, 1967; van Beilen and Poirier, 2007), industrial production of NR depends solely on latex, the cytoplasm of highly specialized cells known as laticifers harvested from the Para rubber tree Hevea brasiliensis. Despite its long history of use as an industrially essential natural material, the molecular mechanisms underlying NR biosynthesis, especially those required for the backbone structure formation, remain to be elucidated.

NR exists in latex as rubber particles (RPs), which primarily consist of a hydrophobic rubber core enclosed by a lipid monolayer (Cornish et al., 1999). In the 1960s, the enzyme responsible for NR biosynthesis from RPs was reported to be rubber transferase (RTase) [EC.2.5.1.20] (Archer, 1969), although its proteinaceous components and catalytic mechanism were not elucidated. The backbone structure of NR comprises of a cis-1,4-polyisoprene with two or three trans-isoprene units at the ω-terminus (Tanaka et al., 1996; Eng et al., 1994) (Figure 1). Generally, the basic backbone structures of cis,trans-mixed polyisoprenoid alcohols, polyprenols and dolichols (C₃₀–C₁₂₀), are biosynthesized via sequential cis-condensation of the C₅ isoprene unit, isopentenyl diphosphate (IPP), onto all-trans short-chain prenyl diphosphates such as geranyl diphosphate (GPP, C₁₀) E,E-farnesyl diphosphate (FPP, C₁₅), and E,E,E-geranylgeranyl diphosphate (GGPP, C₂₀). This reaction is catalyzed by cis-prenyltransferase (cPT) (Takahashi and Koyama, 2006; Koyama, 1999) (Figure 1). The common structural unit of polyisoprenoid alcohols and NR suggests that RTase is a member of the cPT family, although the NR carbon chain length (C_>1500) is much longer than that of other cis,trans-mixed polyisoprenoid alcohols, such as dolichols which are indispensable as sugar carrier lipids during the protein glycosylations in the endoplasmic reticulum (ER) (Helenius and Aebi, 2004).

Figure 1

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We previously isolated two cPT homologues, designated HRT1 and HRT2, from H. brasiliensis, which are predominantly expressed in latex (Asawatreratanakul et al., 2003). Recombinant proteins of HRT1 and HRT2 expressed in Escherichia coli did not exhibit distinct cPT activity independently in vitro. While the recombinant proteins of HRT1 and HRT2, expressed in yeast and Arabidopsis showed significant activity, producing C_80–100 polyisoprenoids, they failed to produce longer chain products corresponding to NR (Takahashi et al., 2012). Recently, cPT homologues expressed in latex of other NR-producing plants, such as Russian dandelion (Taraxacum koksaghyz) (Schmidt et al., 2010a; Schmidt et al., 2010b), lettuce (Lactuca sativa) (Qu et al., 2015) and Euphorbia characias (Spanò et al., 2015), have been isolated, although their recombinant proteins show no RTase activity in vitro. RNA interference-mediated knock-down of cPTs expressed in the latex of Taraxacum brevicorniculatum have been shown to decrease latex rubber content (Post et al., 2012), suggesting the involvement of cPT in NR formation. However, there is no direct evidence to date suggesting that the cPTs expressed in latex are RTases. Therefore, latex-specific cPTs are thought to require eukaryotic cell-specific factor(s) to exhibit cPT activity and additional factor(s) in the latex to modify these cPTs into RTases.

With regard to activation of eukaryotic cPTs, a family of Nogo-B receptor (NgBR) performs the role of regulator of cPT activity. NgBR from humans, originally identified as a receptor for the N-terminal portion of reticulon 4B/Nogo-B (Miao et al., 2006), contains three predicted transmembrane domains in its N-terminal region, and a domain exhibiting low similarity to cPT in the C-terminal half, although it does not have catalytic residues conserved among cPTs. In the cPT-like domain, NgBR can also interact with a human cPT HDS/hCIT (Shridas et al., 2003; Endo et al., 2003) to stabilize it on the ER membrane, resulting in cPT activity enhancement (Harrison et al., 2011). Knock-down (Harrison et al., 2011) or a mutation (Eng et al., 1994) of NgBR causes defects in protein N-glycosylation as a consequence of a reduction in the level of dolichyl phosphates and dolichol-linked oligosaccharides, indicating that NgBR is an HDS-interacting protein and this interaction is critical for its activity. NUS1, a yeast homolog of NgBR, is implicated to have an important role in N-glycosylation and cPT activity (Park et al., 2014). It has also been reported that a defect in LEW1, a counterpart of NgBR in Arabidopsis thaliana, results in reduced levels of dolichol contents and protein glycosylation (Zhang et al., 2008). Despite its importance in the eukaryotic cis,trans-mixed polyisoprenoid biosynthesis, a precise role of NgBR in cPT activity is still unknown owing to a lack of in vitro studies on the function of NgBR family with purified proteins. Moreover, its role as an essential subunit of a deduced cPT-containing protein complex is controversial as various eukaryotic cPTs heterologously expressed in Saccharomyces cerevisiae have shown distinct activity without co-expression of their partner NgBR family protein (Asawatreratanakul et al., 2003; Takahashi et al., 2012; Schmidt et al., 2010b; Shridas et al., 2003; Endo et al., 2003; Harrison et al., 2011; Cunillera et al., 2000; Akhtar et al., 2013; Kera et al., 2012; Surmacz et al., 2014).

In this study, we identified a protein complex that function as the NR biosynthetic machinery on RPs. We identified the partner proteins of HRT1 by RP proteomics and interaction network assays, revealing the formation of a protein complex consisting of HRT1 and two RP-bound proteins, where one of the two was a Hevea NgBR homolog. To investigate functions of the partner proteins, we developed a cell-free translation-coupled recombinant protein introduction system on detergent-washed Hevea RPs (WRPs). This enabled de novo synthesis of NR in vitro by the recombinant protein, providing the first direct evidence that HRT1 is an RTase in H. brasiliensis. In this system, a heterologous cPT from another rubber-producing plant, introduced on Hevea RPs, also exhibited RTase activity comparable to that of HRT1, indicating that accurate introduction of cPT on the WRPs is key to the functional expression of RTase. Reconstitution assays of HRT1 and the partner proteins on WRPs illustrated that the formation of an HRT1-containing ternary complex is important for efficient rubber production via NR biosynthesis from RPs.

In this study, we established a novel system for cell-free, translation-coupled, hydrophobic protein introduction on detergent-washed RPs from H. brasiliensis, allowing de novo synthesis of NR in vitro by the recombinant protein. Additionally, we demonstrated that the key factor for reconstitution of RTase activity of latex cPTs in vitro is proper arrangement of cPT on the specific architecture of WRPs.

Since the first isolation of cPTs, HRT1 and HRT2 from H. brasiliensis latex, as candidates for RTase (Asawatreratanakul et al., 2003), several homologous cDNAs have been isolated from other NR-producing plants, T. koksaghyz (Schmidt et al., 2010a; Schmidt et al., 2010b), L. sativa (Qu et al., 2015) and E. characias (Spanò et al., 2015). However, practical, functional identification of RTase was not accomplished to date, owing to the absence of in vitro RTase activity distinctly exhibited by recombinant cPTs. In a previous study to elucidate activation mechanism of cPT as RTase, we showed that the refolded HRT2, which was heterologously expressed in E. coli and purified from its insoluble membrane fraction under denaturing conditions, was significantly activated by the addition of washed-bottom fraction (BF) of H. brasiliensis latex. This resulted in the formation of polyisoprenes with similar molecular size distributions as that of the NR in Hevea latex (Asawatreratanakul et al., 2003). Whereas the refolded recombinant HRT1 was not activated. BF is the sediment fraction separated by ultracentrifugation of latex and consists mainly of membrane-bound polydisperse particles called lutoids and contaminated RPs (Moir, 1959), with lower RTase activity compared to that of RPs (Tangpakdee et al., 1997; Wititsuwannakul et al., 2003). Therefore, it was suggested that HRT2 might function as a key enzyme in the biosynthesis of natural rubber, correlating with essential co-factor(s) included in the washed-BF. Likewise, eukaryotic long-chain cPTs (Takahashi and Koyama, 2006), responsible for formation of dolichols and/or long-chain (C_>70) polyprenols, such as Rer2p and Srt1p from S. cerevisiae (Sato et al., 1999; Sato et al., 2001), DPS from A. thaliana (Cunillera et al., 2000), and HDS from humans (Endo et al., 2003), do not exhibit cPT activity in vitro when expressed in E. coli. These eukaryotic cPTs were functionally identified by an in vitro assay with crude membrane or microsomal proteins from an yeast mutant strain of RER2 (Sato et al., 1999), expressing a heterologous cPT. This can be described as an almost solo authorised approach for the functional identification to date. In this yeast system, both HRT2 and HRT1 show distinct cPT activity, producing polyisoprenoids with chain-lengths ranging from C₈₀ to C₁₀₀ as general eukaryotic long-chain cPTs. However, their catalytic properties have never been successfully modified into RTase by the addition of any latex fractions (Takahashi et al., 2012). These facts suggest that both, HRT1 and HRT2 may require additional factor(s) present within the eukaryotic cell to exhibit cPT activity and other latex-specific factor(s) to exhibit RTase activity. Additionally, HRT1/HRT2 incorporated into the yeast microsome or membrane fraction may be established as a long-chain cPT even in the presence of latex-derived factor(s). Eukaryotic post-translational modifications such as glycosylations in the ER were considered to be potential candidates for a key activation factor of eukaryotic cPTs generally localised to the ER (Shridas et al., 2003; Sato et al., 2001). However, this is unlikely since in this study, recombinant HRT1 and LsCPT3 showed distinct RTase activity in vitro (Figures 9 and 10) when they were introduced on RPs by means of the wheat germ cell-free system, in which no post-translational modifications occur without supplementation of donor substrates and cofactors (Harbers, 2014).

Figure 10

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In our attempt to explore a candidate protein factor involved in RTase activation, we identified HRBP, an NgBR homologue from H. brasiliensis, as well as HRT1/HRT2, on the detergent-washed RPs (Figure 2—source data 1), while no other enzyme in the natural rubber or IPP biosynthetic pathways was detected. Recently, NgBR homologs on RPs were identified from other NR producing plants, lettuce, and Russian dandelion, using an RP proteomics approach, although no cPT homolog was detected in these proteomics analyses (Qu et al., 2015; Janina, 2015). These NgBR homologs were shown to interact with cPT(s) exclusively expressed in latex. A knockdown of these NgBR homologs results in significant decrease of rubber content, suggesting involvement of the NgBR homologs in the NR biosynthesis of these NR-producing plants. These observations correlate with former reports (Harrison et al., 2011; Park et al., 2014; Zhang et al., 2008; Yu et al., 2006), in which knockdown or mutation of the NgBR family proteins from humans and A. thaliana resulted in severe N-glycosylation defects, owing to decrease in contents of dolichyl phosphates, cis,trans-mixed polyisoprenoids formed by the action of long-chain cPTs. While cPT is encoded by one or two genes in yeasts and animals, in higher plats it is generally encoded by a multigene family, resulting in generation of the short-, middle-, and long-chain cPTs. Short- or middle-chain cPTs from Solanum lycopersicum, heterologously expressed in E. coli, exhibit a distinct cPT activity independently in vitro, producing polyisoprenoids shorter than C₆₀ (Akhtar et al., 2013). Additionally, the recombinant protein of a cis,trans-mixed heptaprenyl diphosphate (C₃₅) synthase from A. thalina, also exhibits a cPT activity independently without corresponding NgBR family protein (Kera et al., 2012; Surmacz et al., 2014). Taken together, the NgBR family protein is considered to be a subunit protein of a long-chain cPT on the ER, responsible for the dolichol or NR formations, resulting in enhancement of cPT activity. However, it is unclear whether involvement of the NgBR family protein is a stringent requirement for cPT activation since various eukaryotic long-chain cPTs, including HRT1 and HRT2, expressed in S. cerevisiae show distinct activity without co-expression of a partner NgBR family protein. In vitro results in the present study emphasize that an NgBR family protein is not necessary for the catalytic activity of cPT to produce polyisoprenoids that correspond to NR; HRT1 and LsCPT3 showed a significant rubber-producing activity when introduced into WRPs (Figures 9, 10). Due to low amino acid sequence identities between HRT1/HRT2, and LsCPT3 and between HRBP and LsCPTL2 (53% and 45%, respectively), it is unlikely that the small amount of HRBP retained on RPs after the detergent wash could have interacted with LsCPT3 as a surrogate partner for its activation. In practice, we have analysed cross-species interaction partnerships between cPT and NgBR family proteins from different plant species by BiFC in the N. benthamiana system, but no distinct interaction between LsCPT3 and HRBP can be detected (unpublished data). It is interesting that coexpression of LsCPT3-LsCPTL2 on WRPs caused a slight increase in efficiency of rubber production, whereas that of HRT1-HRBP did not produce such an effect (Figure 10A). Whether similar phenomena are observed using other cPTs and NgBR homologs from NR-producing plants remains to be determined. Taken together with the observation that distinct fluorescent signals for mTq2-fused HRT1 or HRT2 in N. benthamiana cells were detected only when they were co-expressed with mVanus-fused HRBP (Figure 5), the NgBR family protein may play an important role in proper introduction and folding of cPT or its stabilization on the ER membrane, especially in a heterologous cell expression system, which may be conserved in the RTase in NR-producing plants.

The absence of any RTase activity by HRT1 introduced on liposomes by the cell-free translation-coupled system suggests that structural arrangements of RPs, a hydrophobic core of rubber molecules surrounded by a lipid monolayer, and/or membrane proteins on RPs may be required for HRT1 to exhibit RTase activity. It has been demonstrated that appropriate interactions between lipids and the proteins are required to facilitate accurate folding of membrane proteins during translation (Cymer et al., 2015). To date, various methods for cell-free protein synthesis supplemented with biological membranes or membrane-mimicking structures, such as microsomes, micelles, liposomes, bicelles and nanodiscs, have been developed as a promising strategy to obtain functional membrane proteins. However, it has been pointed out that function and activity of the resulting proteins depend critically on properties of the membrane applied, such as lipid composition, phase, tension, fluidity and curvature (Cymer et al., 2015). Although neither HRT1 nor HRT2 have any deduced transmembrane domains (predicted by TMHMM; http://www.cbs.dtu.dk/services/TMHMM/), these cPTs may require the membrane environment of RPs or the ER for accurate and functional folding, which is considered to correlate with incorporation of HRT1 into WRPs after cell-free synthesis (Figure 7). In our previous study, the refolded HRT2, heterologously expressed in E. coli and purified under a denaturing condition, was significantly activated by the addition of washed BF (Asawatreratanakul et al., 2003). In this case, the partially unfolded HRT2 might be introduced into some membrane systems in BF, microsomal membranes or contaminated RPs, and refolded to exhibit RTase activity. In the present study, we tested a reconstitution of HRT2 in the WRP-based in vitro translation system. However, the protein level was very low compared to that of HRT1 for unknown reason. Also, the WRPs with recombinantly expressed HRT2 did not show distinct RTase activity (data not shown).

Of note was that the size distributions of rubber products from HRT1 and LsCPT3 were quite similar to each other (Figure 10B). These results suggest that the molecular size of de novo synthesised rubber by recombinant cPTs may be determined not by catalytic properties of each cPT but by structural properties of RPs, although further investigations will be needed to elucidate the mechanism for chain-length determination of rubber products. The architecture of RP is considered to be favourable for accommodation of extraordinarily long polyprenyl products formed by sequential condensation of IPP by an RP-localized RTase (Figure 11A). Whereas non-NR-producing eukaryotes also have organelles with a similar structural property, i.e., an ER-derived lipid droplet consists of a hydrophobic core of lipids, triacylglycerols and steryl esters, surrounded by a lipid monolayer (Chrispeels and Herman, 2000). Srt1p was reported to be localized on lipid droplets in S. cerevisiae (Sato et al., 2001), and Nus1p was also detected on lipid droplets using a proteomic approach (Grillitsch et al., 2011), suggesting co-localization or interaction of these proteins on lipid droplets. The fact that yeast does not show RTase activity suggests that arrangement of a cPT with an NgBR family protein on an organelle with a hydrophobic core to accommodate extraordinarily long polyprenyl products may not be a determinant for modification of the cPT into active RTase.

Figure 11

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In spite of high amino acid sequence identity between HRT1 and HRT2 (88%, Figure 2—figure supplement 2), these cPTs showed different subcellular localizations in the N. benthamiana expression system; HRT1 in the Golgi or ER and HRT2 on the plasma membrane (Figure 5A). While subcellular localizations of HRT1 and HRBP are altered from the Golgi to the ER by co-expression of REF, HRT2 is translocated to the plasma membrane, accompanied by HRBP regardless of REF co-expression (Figure 5B), suggesting failure of REF to retain the HRT2-HRBP complex in the ER. These results suggest that cell biological roles of HRT2 in laticifer may be different from those of HRT1. Cell biological roles of cPT on the plasma membrane remain to be elucidated and to the best of our knowledge, this is the first report regarding the identification of a plasma membrane-localised eukaryotic cPT. In addition, HRT1 expression is severely restricted in the latex of H. brasiliensis, while HRT2 transcripts are detected in various tissues (Aoki et al., 2014a). According to the functional characterizations of HRT2 heterologously expressed in E. coli (Asawatreratanakul et al., 2003) and yeast (Takahashi et al., 2012), HRT2 essentially possess catalytic function as cPT or possibly as RTase. Nevertheless, above-mentioned evidences suggest a possibility that HRT1 may be the principal catalytic component of RTase responsible for NR biosynthesis in H. brasiliensis, although we still consider a possible involvement of other cPTs including HRT2 in the NR biosynthesis. Two mechanisms for the NR biosynthesis have been proposed so far (Rojruthai et al., 2010): the first one is de novo formation of NR via sequential cis-1,4-condensation of IPP onto short-chain allylic diphosphates as primer substrates; the second one is elongation or addition of IPP at the α-terminal reactive moiety of pre-exiting rubber molecules or polyprenyl diphosphates in RPs. The allylic diphosphate primer requirement for the RTase reaction (Figure 9C) and no detectable proteins responsible for the short-chain allylic diphosphate biosynthesis on the detergent-washed RPs (Figure 2—source data 1) suggest a possibility that HRT1 on WRP catalyzes a de novo rubber formation using allylic diphosphate as primer substrates. However, an alternative scenario that HRT2 on the WRPs functions as cPT to form polyprenyl diphosphates, which might be utilised by HRT1 as acceptor substrates in the latter NR biosynthetic mechanism, can not be ruled out.

For the next logical step, topologies of HRT1 and HRBP on membrane systems need to be discussed. HRBP is a distantly-related structural homolog of HRT1. The amino acid sequence of HRBP shows some similarity to that of HRT1 (sequence identity: 10.9%), although only HRBP is predicted to have a membrane-spanning region at its N-terminus. As the three-dimensional structure of HRT1 is not yet available, the most suitable templates for predicting the structure of HRBP are bacterial cPTs, which are shown to form homodimers within their crystal structure (Fujihashi et al., 2001). The results from the Y2H and BiFC assays show that HRBP interacts with HRT1, whereas HRT1 does not interact with itself; i.e., HRT1 may form a heteromeric complex with HRBP rather than form a homomultimeric structure on RPs, although HRBP also exhibited self-interaction in the Y2H and BiFC assays. Reconstitution experiments with the two components on WRPs (Figure 9B) support this hypothesis: the reconstituted RTase activity is highest when these two components are expressed. Further experiments are needed to elucidate more precise stoichiometry of the complex.

We demonstrated that HRBP interacted with REF, a well-known RP-bound protein (Berthelot et al., 2014a). The results of this study also revealed the function of REF as a module of the RTase machinery. Although we have no data for the stoichiometry of an HRBP-REF complex, specific interaction partnerships, HRT1-HRBP and HRBP-REF (Figures 3 and 4), and probable formation of the ternary complex (Figure 5B, Figure 5—figure supplement 3) indicate that HRBP functions as a mediating protein between REF and HRT1 to form the ternary complex on RP. Additionally, only when REF was co-expressed with HRT1 and HRBP, the reconstituted RTase activity showed a significant increase. Taken together, the ternary complex including HRT1 may function as rubber-producing machinery on RPs (Figure 11A). Interestingly, the amino acid sequence of SRPP, another RP-abundant protein in H. brasiliensis, contains a REF-homology domain (Oh et al., 1999). To date, several homologous proteins of SRPP have been identified not only from rubber-producing plants but also non-rubber producing plants, designated as the SRPP family or stress-related protein family (Berthelot et al., 2014a). However, no counterpart of REF, a shorter protein than SRPP family, has been found from other species. Laibach et al. (2015) reported that silencing of a REF-related gene TbREF affected the rubber production in the RP of Russian dandelion, even though this gene is rather homologous to SRPP. It is unknown whether TbREF interacts with other component(s) such as an NgBR homologue of Russian dandelion. The system for in vitro translation-coupled introduction of hydrophobic proteins on RPs, developed in this study, also enables us to shed light on molecular function of REF and SRPP on RPs. Our observations of REF in modulating diameters of WRPs (Figure 8) represents the first direct evidence of REF being an important protein in the WRP stability. Berthelot et al. (2014b) reported different behaviors of REF and SRPP on model membrane systems, where REF inserts into membranes and disrupts their integrity, whereas SRPP weakly interacts with membranes without any disruption. Since REF is likely to insert itself into RP, it is expected to interact with rubber molecules in RP. Our future plans include study of whether REF controls the turnover of RTase by interacting with the rubber molecule produced by HRT1 in a deeper region of the RP membrane.

Similarities between the structures of RP and the ER-derived lipid droplet (i.e. a hydrophobic core surrounded by a lipid monolayer) suggest that RPs may originate from the ER or Golgi apparatus (Chrispeels and Herman, 2000). This speculation was bolstered by identification of various proteins homologous to vesicular trafficking-related proteins in the ER or Golgi apparatus, on WRPs (Figure 2—source data 1). For example, Rab GTPases, RabB (ER to Golgi), RabD (ER to Golgi) and RabH (the Golgi), a Sar1-type coat GTPase (ER to Golgi), a YKT6 type R-SNARE (trans-Golgi network, TGN), and clathrins (TGN) (Nielsen et al., 2008; Bassham et al., 2008). However, the co-existence of marker proteins for other vesicles involved in different phases of membrane trafficking pathways, e.g. the endocytic/endosomal compartments (RabA, RabC, RabE, RabF, an ArfB-type coat GTPase and clathrins), vacuoles (RabG) and lipid-bodies (β-1,3-glucase [Paul et al., 2014]), on RPs suggests that RP formation and maturation may be complicated by fusion of multiple vesicles. Different subcellular localization of the protein complexes HRT1–HRBP and HRBP–REF may partly reflect the stepwise formation of the HRT1–HRBP–REF complex on RPs (Figure 11B). In addition, proteomics of the detergent-washed RPs did not identify any major lipid droplet proteins such as seipin, perilipins or oleosins (Murphy, 2012), consistent with a previous proteomics study of RPs (Dai et al., 2013). This implies that mechanisms for de novo formation from the ER and maturation of RPs may be different from those of lipid droplets.

In conclusion, this study represents important progress toward understanding the molecular machinery of rubber production on RP. Identification of the two partner proteins of HRT1 provides suggestions for possible formation of a ternary complex that functions as the RTase. Although the interactions between cPTs and NgBR homologs are generally assumed in other organisms, HRBP-REF interaction discovered in this study is specific to NR production. Cell biological analyses of the localization of the three components suggest that RPs can be generated by a fusion of various cellular vesicles. We demonstrated in vitro rubber production on WRP for the first time by heterologously expressing cPTs from the Para rubber tree and lettuce, although the assay system developed in this study still included a small amount of endogenous proteins sustained on WRPs. We also showed that HRBP and REF promoted efficient rubber synthesis in the same system when these two proteins were co-expressed with HRT1. Future work will include investigating the structures and functions of the two partner proteins of HRT1. In particular, the effect of REF on the alterations of specific membrane systems is a noteworthy observation. The in vitro translation-coupled protein reconstitution on WRPs is a versatile system for total elucidation of NR biosynthesis and may lead to molecular breeding of transgenic plants with enhanced NR biosynthetic activity.

1. 1. Takahashi S
   2. Nakayama T

   (2015) Hevea brasiliensis HRTBP mRNA for transferase binding protein

   Publicly available at the DNA Data Bank of Japan (accession no: LC057267).

   http://getentry.ddbj.nig.ac.jp/getentry/na/LC057267/?format=flatfile&filetype=html&trace=true&show_suppressed=false&limit=10

1. 1. Qu Y
   2. Chakrabarty R
   3. Tran HT
   4. Kwon EJ
   5. Kwon M
   6. Nguyen TD
   7. Ro DK

   (2015) Lactuca sativa cis-prenyltransferase 3 mRNA, complete cds.

   Publicly available at the NCBI Nucleotide (accession no: F752488).

   http://www.ncbi.nlm.nih.gov/nuccore/KF752488
2. 1. Rahman AY
   2. Usharraj AO
   3. Misra BB
   4. Thottathil GP
   5. Jayasekaran K
   6. Feng Y
   7. Hou S
   8. Ong SY
   9. Ng FL
   10. Lee LS
   11. Tan HS
   12. Sakaff MK
   13. Teh BS
   14. Khoo BF
   15. Badai SS
   16. Aziz NA
   17. Yuryev A
   18. Knudsen B
   19. Dionne-Laporte A
   20. McHunu NP
   21. Yu Q
   22. Langston BJ
   23. Freitas TA
   24. Young AG
   25. Chen R
   26. Wang L
   27. Najimudin N
   28. Saito JA
   29. Alam M

   (2013) TSA: Hevea brasiliensis contigxxxxx, mRNA sequence.

   Publicly available at the NCBI BioProject (accession no: PRJNA828950).

   https://www.ncbi.nlm.nih.gov/nuccore/?term=PRJNA82895
3. 1. Qu Y
   2. Chakrabarty R
   3. Tran HT
   4. Kwon EJ
   5. Kwon M
   6. Nguyen TD
   7. Ro DK

   (2016) Lactuca sativa cis-prenyltransferase-like 2 mRNA, complete cds

   Publicly available at the NCBI Nucleotide (accession no: KF752485).

   http://www.ncbi.nlm.nih.gov/nuccore/687843842/

1. 1. Akhtar TA
   2. Matsuba Y
   3. Schauvinhold I
   4. Yu G
   5. Lees HA
   6. Klein SE
   7. Pichersky E

   (2013) The tomato cis-prenyltransferase gene family

   The Plant Journal 73:640–652.

   https://doi.org/10.1111/tpj.12063

   • PubMed
   • Google Scholar
2. 1. Aoki Y
   2. Takahashi S
   3. Takayama D
   4. Ogata Y
   5. Sakurai N
   6. Suzuki H
   7. Asawatreratanakul K
   8. Wititsuwannakul D
   9. Wititsuwannakul R
   10. Shibata D
   11. Koyama T
   12. Nakayama T

   (2014a) Identification of laticifer-specific genes and their promoter regions from a natural rubber producing plant Hevea brasiliensis

   Plant Science 225:1–8.

   https://doi.org/10.1016/j.plantsci.2014.05.003

   • PubMed
   • Google Scholar
3. 1. Aoki Y
   2. Takahashi S
   3. Toda S
   4. Koyama T
   5. Nakayama T

   (2014b) Transcriptional responses of laticifer-specific genes to phytohormones in a suspension-cultured cell line derived from petioles of Hevea brasiliensis

   Plant Biotechnology 31:593–598.

   https://doi.org/10.5511/plantbiotechnology.14.1015a

   • Google Scholar
4. Book

   1. Archer BL
   2. Cockbain EG

   (1969)

   Methods Enzymol

   Clayton Raymond B, editors. Academic Press.

   • Google Scholar
5. 1. Asawatreratanakul K
   2. Zhang YW
   3. Wititsuwannakul D
   4. Wititsuwannakul R
   5. Takahashi S
   6. Rattanapittayaporn A
   7. Koyama T

   (2003) Molecular cloning, expression and characterization of cDNA encoding cis-prenyltransferases from Hevea brasiliensis. A key factor participating in natural rubber biosynthesis

   European Journal of Biochemistry 270:4671–4680.

   https://doi.org/10.1046/j.1432-1033.2003.03863.x

   • PubMed
   • Google Scholar
6. 1. Bassham DC
   2. Brandizzi F
   3. Otegui MS
   4. Sanderfoot AA

   (2008) The secretory system of Arabidopsis

   The Arabidopsis Book 6:e0116.

   https://doi.org/10.1199/tab.0116

   • PubMed
   • Google Scholar
7. 1. Berthelot K
   2. Lecomte S
   3. Estevez Y
   4. Peruch F

   (2014a) Hevea brasiliensis REF (Hev b 1) and SRPP (Hev b 3): An overview on rubber particle proteins

   Biochimie 106:1–9.

   https://doi.org/10.1016/j.biochi.2014.07.002

   • PubMed
   • Google Scholar
8. 1. Berthelot K
   2. Lecomte S
   3. Estevez Y
   4. Zhendre V
   5. Henry S
   6. Thévenot J
   7. Dufourc EJ
   8. Alves ID
   9. Peruch F

   (2014b) Rubber particle proteins, HbREF and HbSRPP, show different interactions with model membranes

   Biochimica Et Biophysica Acta (BBA) - Biomembranes 1838:287–299.

   https://doi.org/10.1016/j.bbamem.2013.08.025

   • Google Scholar
9. 1. Brasher MI
   2. Surmacz L
   3. Leong B
   4. Pitcher J
   5. Swiezewska E
   6. Pichersky E
   7. Akhtar TA

   (2015) A two-component enzyme complex is required for dolichol biosynthesis in tomato

   The Plant Journal 82:903–914.

   https://doi.org/10.1111/tpj.12859

   • PubMed
   • Google Scholar
10. 1. Chow KS
    2. Wan KL
    3. Isa MN
    4. Bahari A
    5. Tan SH
    6. Harikrishna K
    7. Yeang HY

    (2007) Insights into rubber biosynthesis from transcriptome analysis of Hevea brasiliensis latex

    Journal of Experimental Botany 58:2429–2440.

    https://doi.org/10.1093/jxb/erm093

    • PubMed
    • Google Scholar
11. 1. Chrispeels MJ
    2. Herman EM

    (2000) Endoplasmic reticulum-derived compartments function in storage and as mediators of vacuolar remodeling via a new type of organelle, precursor protease vesicles

    Plant Physiology 123:1227–1234.

    https://doi.org/10.1104/pp.123.4.1227

    • PubMed
    • Google Scholar
12. 1. Cornish K
    2. Wood DF
    3. Windle JJ

    (1999) Rubber particles from four different species, examined by transmission electron microscopy and electron-paramagnetic-resonance spin labeling, are found to consist of a homogeneous rubber core enclosed by a contiguous, monolayer biomembrane

    Planta 210:85–96.

    https://doi.org/10.1007/s004250050657

    • PubMed
    • Google Scholar
13. 1. Cunillera N
    2. Arró M
    3. Forés O
    4. Manzano D
    5. Ferrer A

    (2000) Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis

    FEBS Letters 477:170–174.

    https://doi.org/10.1016/s0014-5793(00)01798-1

    • PubMed
    • Google Scholar
14. 1. Cymer F
    2. von Heijne G
    3. White SH

    (2015) Mechanisms of integral membrane protein insertion and folding

    Journal of Molecular Biology 427:999–1022.

    https://doi.org/10.1016/j.jmb.2014.09.014

    • PubMed
    • Google Scholar
15. 1. Dai L
    2. Kang G
    3. Li Y
    4. Nie Z
    5. Duan C
    6. Zeng R

    (2013) In-depth proteome analysis of the rubber particle of Hevea brasiliensis (para rubber tree)

    Plant Molecular Biology 82:155–168.

    https://doi.org/10.1007/s11103-013-0047-y

    • PubMed
    • Google Scholar
16. 1. Dennis MS
    2. Light DR

    (1989)

    Rubber elongation factor from Hevea brasiliensis. Identification, characterization, and role in rubber biosynthesis

    The Journal of Biological Chemistry 264:18608–18617.

    • PubMed
    • Google Scholar
17. 1. Endo S
    2. Zhang Y-W
    3. Takahashi S
    4. Koyama T

    (2003) Identification of human dehydrodolichyl diphosphate synthase gene

    Biochimica Et Biophysica Acta (BBA) - Gene Structure and Expression 1625:291–295.

    https://doi.org/10.1016/S0167-4781(02)00628-0

    • Google Scholar
18. 1. Eng AH
    2. Kawahara S
    3. Tanaka Y

    (1994) Trans-isoprene units in natural rubber

    Rubber Chemistry and Technology 67:159–168.

    https://doi.org/10.5254/1.3538662

    • Google Scholar
19. 1. Fujihashi M
    2. Zhang Y-W
    3. Higuchi Y
    4. Li X-Y
    5. Koyama T
    6. Miki K

    (2001) Crystal structure of cis-prenyl chain elongating enzyme, undecaprenyl diphosphate synthase

    PNAS 98:4337–4342.

    https://doi.org/10.1073/pnas.071514398

    • Google Scholar
20. 1. Gookin TE
    2. Assmann SM

    (2014) Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors

    Plant Journal 80:553–567.

    https://doi.org/10.1111/tpj.12639

    • PubMed
    • Google Scholar
21. 1. Grillitsch K
    2. Connerth M
    3. Köfeler H
    4. Arrey TN
    5. Rietschel B
    6. Wagner B
    7. Karas M
    8. Daum G

    (2011) Lipid particles/droplets of the yeast Saccharomyces cerevisiae revisited: Lipidome meets Proteome

    Biochimica Et Biophysica Acta 1811:1165–1176.

    https://doi.org/10.1016/j.bbalip.2011.07.015

    • Google Scholar
22. 1. Han KH
    2. Shin DH
    3. Yang J
    4. Kim IJ
    5. Oh SK
    6. Chow KS

    (2000) Genes expressed in the latex of Hevea brasiliensis

    Tree Physiology 20:503–510.

    https://doi.org/10.1093/treephys/20.8.503

    • PubMed
    • Google Scholar
23. 1. Harbers M

    (2014) Wheat germ systems for cell-free protein expression

    FEBS Letters 588:2762–2773.

    https://doi.org/10.1016/j.febslet.2014.05.061

    • PubMed
    • Google Scholar
24. 1. Harrison KD
    2. Park EJ
    3. Gao N
    4. Kuo A
    5. Rush JS
    6. Waechter CJ
    7. Lehrman MA
    8. Sessa WC

    (2011) Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation

    The EMBO Journal 30:2490–2500.

    https://doi.org/10.1038/emboj.2011.147

    • PubMed
    • Google Scholar
25. 1. Helenius A
    2. Aebi M

    (2004) Roles of N-linked glycans in the endoplasmic reticulum

    Annual Review of Biochemistry 73:1019–1049.

    https://doi.org/10.1146/annurev.biochem.73.011303.073752

    • PubMed
    • Google Scholar
26. 1. Janina E

    (2015) A rubber transferase activator is necessary for natural rubber biosynthesis in dandelion

     Nature Plants 1:15048.

    https://doi.org/10.1038/nplants.2015.48

    • Google Scholar
27. 1. Kera K
    2. Takahashi S
    3. Sutoh T
    4. Koyama T
    5. Nakayama T

    (2012) Identification and characterization of a cis,trans-mixed heptaprenyl diphosphate synthase from Arabidopsis thaliana

    The FEBS Journal 279:3813–3827.

    https://doi.org/10.1111/j.1742-4658.2012.08742.x

    • PubMed
    • Google Scholar
28. 1. Ko JH
    2. Chow KS
    3. Han KH

    (2003) Transcriptome analysis reveals novel features of the molecular events occurring in the laticifers of Hevea brasiliensis (para rubber tree)

    Plant Molecular Biology 53:479–492.

    https://doi.org/10.1023/B:PLAN.0000019119.66643.5d

    • PubMed
    • Google Scholar
29. 1. Koyama T

    (1999) Molecular analysis of prenyl chain elongating enzymes

    Bioscience, Biotechnology, and Biochemistry 63:1671–1676.

    https://doi.org/10.1271/bbb.63.1671

    • PubMed
    • Google Scholar
30. 1. Kush A
    2. Goyvaerts E
    3. Chye ML
    4. Chua NH

    (1990) Laticifer-specific gene expression in Hevea brasiliensis (rubber tree)

    PNAS 87:1787–1790.

    https://doi.org/10.1073/pnas.87.5.1787

    • Google Scholar
31. 1. Laibach N
    2. Hillebrand A
    3. Twyman RM
    4. Prüfer D
    5. Schulze Gronover C

    (2015) Identification of a Taraxacum brevicorniculatum rubber elongation factor protein that is localized on rubber particles and promotes rubber biosynthesis

    The Plant Journal 82:609–620.

    https://doi.org/10.1111/tpj.12836

    • PubMed
    • Google Scholar
32. 1. Metcalfe CR

    (1967) Distribution of latex in the plant kingdom

    Economic Botany 21:115–127.

    https://doi.org/10.1007/BF02897859

    • Google Scholar
33. 1. Miao RQ
    2. Gao Y
    3. Harrison KD
    4. Prendergast J
    5. Acevedo LM
    6. Yu J
    7. Hu F
    8. Strittmatter SM
    9. Sessa WC

    (2006) Identification of a receptor necessary for Nogo-B stimulated chemotaxis and morphogenesis of endothelial cells

    PNAS 103:10997–11002.

    https://doi.org/10.1073/pnas.0602427103

    • Google Scholar
34. 1. Moir GFJ

    (1959) Ultracentrifugation and Staining of Hevea Latex

    Nature 184:1626–1628.

    https://doi.org/10.1038/1841626a0

    • Google Scholar
35. 1. Murphy DJ

    (2012) The dynamic roles of intracellular lipid droplets: from archaea to mammals

    Protoplasma 249:541–585.

    https://doi.org/10.1007/s00709-011-0329-7

    • PubMed
    • Google Scholar
36. 1. Nelson BK
    2. Cai X
    3. Nebenführ A

    (2007) A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants

    The Plant Journal 51:1126–1136.

    https://doi.org/10.1111/j.1365-313X.2007.03212.x

    • PubMed
    • Google Scholar
37. 1. Nielsen E
    2. Cheung AY
    3. Ueda T

    (2008) The regulatory RAB and ARF GTPases for vesicular trafficking

    Plant Physiology 147:1516–1526.

    https://doi.org/10.1104/pp.108.121798

    • PubMed
    • Google Scholar
38. 1. Nozawa A
    2. Nanamiya H
    3. Tozawa Y

    (2010) Production of membrane proteins through the wheat-germ cell-free technology

    Methods in Molecular Biology 607:213–218.

    https://doi.org/10.1007/978-1-60327-331-2_17

    • PubMed
    • Google Scholar
39. 1. Oh SK
    2. Kang H
    3. Shin DH
    4. Yang J
    5. Chow KS
    6. Yeang HY
    7. Wagner B
    8. Breiteneder H
    9. Han KH

    (1999) Isolation, characterization, and functional analysis of a novel cDNA clone encoding a small rubber particle protein from Hevea brasiliensis

    Journal of Biological Chemistry 274:17132–17138.

    https://doi.org/10.1074/jbc.274.24.17132

    • PubMed
    • Google Scholar
40. 1. Pagny S
    2. Bouissonnie F
    3. Sarkar M
    4. Follet-Gueye ML
    5. Driouich A
    6. Schachter H
    7. Faye L
    8. Gomord V

    (2003) Structural requirements for Arabidopsis beta1,2-xylosyltransferase activity and targeting to the Golgi

    The Plant Journal 33:189–203.

    https://doi.org/10.1046/j.0960-7412.2002.01604.x

    • PubMed
    • Google Scholar
41. 1. Palmiter RD

    (1974) Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid

    Biochemistry 13:3606–3615.

    https://doi.org/10.1021/bi00714a032

    • PubMed
    • Google Scholar
42. 1. Park EJ
    2. Grabińska KA
    3. Guan Z
    4. Stránecký V
    5. Hartmannová H
    6. Hodaňová K
    7. Barešová V
    8. Sovová J
    9. Jozsef L
    10. Ondrušková N
    11. Hansíková H
    12. Honzík T
    13. Zeman J
    14. Hůlková H
    15. Wen R
    16. Kmoch S
    17. Sessa WC

    (2014) Mutation of Nogo-B receptor, a subunit of cis-prenyltransferase, causes a congenital disorder of glycosylation

    Cell Metabolism 20:448–457.

    https://doi.org/10.1016/j.cmet.2014.06.016

    • PubMed
    • Google Scholar
43. 1. Paul LK
    2. Rinne PLH
    3. van der Schoot C

    (2014) Refurbishing the plasmodesmal chamber: a role for lipid bodies?

    Frontiers in Plant Science 5:40.

    https://doi.org/10.3389/fpls.2014.00040

    • Google Scholar
44. 1. Post J
    2. van Deenen N
    3. Fricke J
    4. Kowalski N
    5. Wurbs D
    6. Schaller H
    7. Eisenreich W
    8. Huber C
    9. Twyman RM
    10. Prüfer D
    11. Gronover CS

    (2012) Laticifer-specific cis-prenyltransferase silencing affects the rubber, triterpene, and inulin content of Taraxacum brevicorniculatum

    Plant Physiology 158:1406–1417.

    https://doi.org/10.1104/pp.111.187880

    • PubMed
    • Google Scholar
45. 1. Qu Y
    2. Chakrabarty R
    3. Tran HT
    4. Kwon EJ
    5. Kwon M
    6. Nguyen TD
    7. Ro DK

    (2015) A lettuce (Lactuca sativa) homolog of human Nogo-B receptor interacts with cis-prenyltransferase and is necessary for natural rubber biosynthesis

    The Journal of Biological Chemistry 290:1898–1914.

    https://doi.org/10.1074/jbc.M114.616920

    • PubMed
    • Google Scholar
46. 1. Rahman AY
    2. Usharraj AO
    3. Misra BB
    4. Thottathil GP
    5. Jayasekaran K
    6. Feng Y
    7. Hou S
    8. Ong SY
    9. Ng FL
    10. Lee LS
    11. Tan HS
    12. Sakaff MK
    13. Teh BS
    14. Khoo BF
    15. Badai SS
    16. Aziz NA
    17. Yuryev A
    18. Knudsen B
    19. Dionne-Laporte A
    20. Mchunu NP
    21. Yu Q
    22. Langston BJ
    23. Freitas TA
    24. Young AG
    25. Chen R
    26. Wang L
    27. Najimudin N
    28. Saito JA
    29. Alam M

    (2013) Draft genome sequence of the rubber tree Hevea brasiliensis

    BMC Genomics 14:75.

    https://doi.org/10.1186/1471-2164-14-75

    • PubMed
    • Google Scholar
47. 1. Rojruthai P
    2. Sakdapipanich JT
    3. Takahashi S
    4. Hyegin L
    5. Noike M
    6. Koyama T
    7. Tanaka Y

    (2010) In vitro synthesis of high molecular weight rubber by Hevea small rubber particles

    Journal of Bioscience and Bioengineering 109:107–114.

    https://doi.org/10.1016/j.jbiosc.2009.08.009

    • PubMed
    • Google Scholar
48. 1. Saint-Jore-Dupas C
    2. Nebenfuhr A
    3. Boulaflous A
    4. Follet-Gueye M-L
    5. Plasson C
    6. Hawes C
    7. Driouich A
    8. Faye L
    9. Gomord V

    (2006) Plant N-Glycan processing enzymes employ different targeting mechanisms for their spatial arrangement along the secretory pathway

    The Pland Cell 18:3182–3200.

    https://doi.org/10.1105/tpc.105.036400

    • Google Scholar
49. 1. Sato M
    2. Fujisaki S
    3. Sato K
    4. Nishimura Y
    5. Nakano A

    (2001) Yeast Saccharomyces cerevisiae has two cis-prenyltransferases with different properties and localizations. Implication for their distinct physiological roles indolichol synthesis

    Genes to Cells 6:495–506.

    https://doi.org/10.1046/j.1365-2443.2001.00438.x

    • PubMed
    • Google Scholar
50. 1. Sato M
    2. Sato K
    3. Nishikawa S
    4. Hirata A
    5. Kato J
    6. Nakano A

    (1999) The yeast RER2 gene, identified by endoplasmic reticulum protein localization mutations, encodes cis-prenyltransferase, a key enzyme in dolichol synthesis

    Molecular and Cellular Biology 19:471–483.

    https://doi.org/10.1128/mcb.19.1.471

    • PubMed
    • Google Scholar
51. 1. Schmidt T
    2. Hillebrand A
    3. Wurbs D
    4. Wahler D
    5. Lenders M
    6. Schulze Gronover C
    7. Prüfer D

    (2010a) Molecular cloning and characterization of rubber biosynthetic genes from Taraxacum koksaghyz

    Plant Molecular Biology Reporter 28:277–284.

    https://doi.org/10.1007/s11105-009-0145-9

    • Google Scholar
52. 1. Schmidt T
    2. Lenders M
    3. Hillebrand A
    4. van Deenen N
    5. Munt O
    6. Reichelt R
    7. Eisenreich W
    8. Fischer R
    9. Prüfer D
    10. Gronover CS

    (2010b) Characterization of rubber particles and rubber chain elongation in Taraxacum koksaghyz

    BMC Biochemistry 11:11.

    https://doi.org/10.1186/1471-2091-11-11

    • PubMed
    • Google Scholar
53. 1. Shridas P
    2. Rush JS
    3. Waechter CJ

    (2003) Identification and characterization of a cDNA encoding a long-chain cis-isoprenyltranferase involved in dolichyl monophosphate biosynthesis in the ER of brain cells

    Biochemical and Biophysical Research Communications 312:1349–1356.

    https://doi.org/10.1016/j.bbrc.2003.11.065

    • PubMed
    • Google Scholar
54. 1. Spanò D
    2. Pintus F
    3. Esposito F
    4. Loche D
    5. Floris G
    6. Medda R

    (2015) Euphorbia characias latex: micromorphology of rubber particles and rubber transferase activity

    Plant Physiology and Biochemistry 87:26–34.

    https://doi.org/10.1016/j.plaphy.2014.12.008

    • PubMed
    • Google Scholar
55. 1. Surmacz L
    2. Plochocka D
    3. Kania M
    4. Danikiewicz W
    5. Swiezewska E

    (2014) cis-Prenyltransferase AtCPT6 produces a family of very short-chain polyisoprenoids in planta

    Biochimica Et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids 1841:240–250.

    https://doi.org/10.1016/j.bbalip.2013.11.011

    • Google Scholar
56. 1. Takahashi S
    2. Koyama T

    (2006) Structure and function of cis-prenyl chain elongating enzymes

    The Chemical Record 6:194–205.

    https://doi.org/10.1002/tcr.20083

    • PubMed
    • Google Scholar
57. 1. Takahashi S
    2. Lee H-J
    3. Yamashita S
    4. Koyama T

    (2012) Characterization of cis-prenyltransferases from the rubber producing plant Hevea brasiliensis heterologously expressed in yeast and plant cells

    Plant Biotechnology 29:411–417.

    https://doi.org/10.5511/plantbiotechnology.12.0625a

    • Google Scholar
58. 1. Tanaka Y
    2. Aik-Hwee E
    3. Ohya N
    4. Nishiyama N
    5. Tangpakdee J
    6. Kawahara S
    7. Wititsuwannakul R

    (1996) Initiation of rubber biosynthesis in Hevea brasiliensis: characterization of initiating species by structural analysis

    Phytochemistry 41:1501–1505.

    https://doi.org/10.1016/0031-9422(95)00817-9

    • Google Scholar
59. 1. Tangpakdee J
    2. Tanaka Y
    3. Ogura K
    4. Koyama T
    5. Wititsuwannakul R
    6. Wititsuwannakul D

    (1997) Rubber formation by fresh bottom fraction of hevea latex

    Phytochemistry 45:269–274.

    https://doi.org/10.1016/S0031-9422(96)00838-2

    • Google Scholar
60. 1. van Beilen JB
    2. Poirier Y

    (2007) Establishment of new crops for the production of natural rubber

    Trends in Biotechnology 25:522–529.

    https://doi.org/10.1016/j.tibtech.2007.08.009

    • PubMed
    • Google Scholar
61. 1. Wititsuwannakul D
    2. Rattanapittayaporn A
    3. Wititsuwannakul R

    (2003) Rubber biosynthesis by aHevea latex bottom-fraction membrane

    Journal of Applied Polymer Science 87:90–96.

    https://doi.org/10.1002/app.11674

    • Google Scholar
62. 1. Xiang Q
    2. Xia K
    3. Dai L
    4. Kang G
    5. Li Y
    6. Nie Z
    7. Duan C
    8. Zeng R

    (2012) Proteome analysis of the large and the small rubber particles of Hevea brasiliensis using 2D-DIGE

    Plant Physiology and Biochemistry 60:207–213.

    https://doi.org/10.1016/j.plaphy.2012.08.010

    • PubMed
    • Google Scholar
63. 1. Yu L
    2. Peña Castillo L
    3. Mnaimneh S
    4. Hughes TR
    5. Brown GW
    6. Castillo LP

    (2006) A survey of essential gene function in the yeast cell division cycle

    Molecular Biology of the Cell 17:4736–4747.

    https://doi.org/10.1091/mbc.E06-04-0368

    • PubMed
    • Google Scholar
64. 1. Zhang H
    2. Ohyama K
    3. Boudet J
    4. Chen Z
    5. Yang J
    6. Zhang M
    7. Muranaka T
    8. Maurel C
    9. Zhu JK
    10. Gong Z

    (2008) Dolichol biosynthesis and its effects on the unfolded protein response and abiotic stress resistance in Arabidopsis

    The Pland Cell 20:1879–1898.

    https://doi.org/10.1105/tpc.108.061150

    • PubMed
    • Google Scholar

Author details

1. Satoshi Yamashita

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Present address

   Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa, Japan

   Contribution

   SY, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article

   Competing interests

   The authors declare that no competing interests exist.

2. Haruhiko Yamaguchi

   Sumitomo Rubber Industries, Ltd, Kobe, Japan

   Contribution

   HY, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article, Contributed unpublished essential data or reagents

   Competing interests

   The authors declare that no competing interests exist.

3. Toshiyuki Waki

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Contribution

   TW, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article, Contributed unpublished essential data or reagents

   Competing interests

   The authors declare that no competing interests exist.

4. Yuichi Aoki

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Present address

   Graduate School of Information Sciences, Tohoku University, Sendai, Japan

   Contribution

   YA, Acquisition of data, Analysis and interpretation of data

   Competing interests

   The authors declare that no competing interests exist.

5. Makie Mizuno

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Contribution

   MM, Acquisition of data, Analysis and interpretation of data

   Competing interests

   The authors declare that no competing interests exist.

6. Fumihiro Yanbe

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Contribution

   FY, Acquisition of data, Analysis and interpretation of data

   Competing interests

   The authors declare that no competing interests exist.

7. Tomoki Ishii

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Contribution

   TI, Acquisition of data, Analysis and interpretation of data

   Competing interests

   The authors declare that no competing interests exist.

8. Ayuta Funaki

   Graduate School of Engineering, Tohoku University, Sendai, Japan

   Contribution

   AF, Acquisition of data, Analysis and interpretation of data

   Competing interests

   The authors declare that no competing interests exist.

9. Yuzuru Tozawa

   Graduate School of Science and Engineering, Saitama University, Saitama, Japan

   Contribution

   YT, Conception and design, Analysis and interpretation of data, Drafting or revising the article, Contributed unpublished essential data or reagents

   Competing interests

   The authors declare that no competing interests exist.

10. Yukino Miyagi-Inoue

    Sumitomo Rubber Industries, Ltd, Kobe, Japan

    Contribution

    YM-I, Acquisition of data, Analysis and interpretation of data, Contributed unpublished essential data or reagents

    Competing interests

    The authors declare that no competing interests exist.

11. Kazuhisa Fushihara

    Sumitomo Rubber Industries, Ltd, Kobe, Japan

    Contribution

    KF, Conception and design, Analysis and interpretation of data, Contributed unpublished essential data or reagents

    Competing interests

    The authors declare that no competing interests exist.

12. Toru Nakayama

    Graduate School of Engineering, Tohoku University, Sendai, Japan

    Contribution

    TN, Conception and design, Analysis and interpretation of data, Drafting or revising the article

    Competing interests

    The authors declare that no competing interests exist.

13. Seiji Takahashi

    Graduate School of Engineering, Tohoku University, Sendai, Japan

    Contribution

    ST, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article, Contributed unpublished essential data or reagents

    For correspondence

    takahasi@seika.che.tohoku.ac.jp

    Competing interests

    The authors declare that no competing interests exist.

    "This ORCID iD identifies the author of this article:" 0000-0002-2288-4340

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