The ER membrane protein complex interacts cotranslationally to enable biogenesis of multipass membrane proteins
- University of California, San Francisco, United States
- Max Planck Institute of Biochemistry, Germany
- University of Pittsburgh, United States
Abstract
The endoplasmic reticulum (ER) supports biosynthesis of proteins with diverse transmembrane domain (TMD) lengths and hydrophobicity. Features in transmembrane domains such as charged residues in ion channels are often functionally important, but could pose a challenge during cotranslational membrane insertion and folding. Our systematic proteomic approaches in both yeast and human cells revealed that the ER membrane protein complex (EMC) binds to and promotes the biogenesis of a range of multipass transmembrane proteins, with a particular enrichment for transporters. Proximity-specific ribosome profiling demonstrates that the EMC engages clients cotranslationally and immediately following clusters of TMDs enriched for charged residues. The EMC can remain associated after completion of translation, which both protects clients from premature degradation and allows recruitment of substrate-specific and general chaperones. Thus, the EMC broadly enables the biogenesis of multipass transmembrane proteins containing destabilizing features, thereby mitigating the trade-off between function and stability.
Data availability
Sequencing data have been deposited in GEO under accession code GSE112891.
-
The ER membrane protein complex interacts cotranslationally to enable biogenesis of multipass membrane proteinsPublicly available at the NCBI Gene Expression Omnibus (accession no: GSE112891).
-
The SND proteins constitute an alternative targeting route to the endoplasmic reticulumPublicly available at the NCBI Gene Expression Omnibus (accession no: GSE85686).
-
Principles of ER Co-Translational Translocation Revealed by Proximity-Specific Ribosome ProfilingPublicly available at the NCBI Gene Expression Omnibus (accession no: GSE61012).
Article and author information
Author details
Jonathan S Weissman
Funding
Howard Hughes Medical Institute (Investigator Program)
- Jonathan S Weissman
Deutsche Forschungsgemeinschaft (Gottfried Wilhelm Leibniz Prize MA 1764/2-1)
- Georg HH Borner
European Research Council (ERC2012-SyG_318987-ToPAG)
- Daniel N Itzhak
Howard Hughes Medical Institute (Faculty Scholar Grant)
- Adam Frost
National Institutes of Health (AG041826)
- Jonathan S Weissman
National Institutes of Health (1DP2GM110772-01)
- Adam Frost
National Institutes of Health (8P41GM103481)
- Alma L Burlingame
National Institutes of Health (1S10OD16229)
- Alma L Burlingame
National Institutes of Health (GM075061)
- Jeffrey Brodsky
Helen Hay Whitney Foundation (Postdoctoral Fellowship)
- Matthew J Shurtleff
Jane Coffin Childs Memorial Fund for Medical Research (Postdoctoral Fellowship)
- Nicole T Schirle Oakdale
Sandler Foundation (Program for Breakthrough Biomedical Research)
- Adam Frost
American Asthma Foundation
- Adam Frost
Louis-Jeantet Foundation
- Daniel N Itzhak
Dr. Miriam and Sheldon G. Adelson Medical Research Foundation
- Alma L Burlingame
Max Planck Society for the Advancement of Science
- Daniel N Itzhak
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- David Ron, University of Cambridge, United Kingdom
Version history
- Received: March 27, 2018
- Accepted: May 26, 2018
- Accepted Manuscript published: May 29, 2018 (version 1)
- Version of Record published: June 11, 2018 (version 2)
- Version of Record updated: August 6, 2018 (version 3)
Copyright
© 2018, Shurtleff et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 13,673
- views
-
- 1,752
- downloads
-
- 170
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
The ER membrane protein complex interacts cotranslationally to enable biogenesis of multipass membrane proteins
eLife 7:e37018.
https://doi.org/10.7554/eLife.37018
Further reading
-
- Cancer Biology
- Cell Biology
Establishing a zebrafish model of a deadly type of brain tumor highlights the role of the immune system in the early stages of the disease.
-
- Cell Biology
- Neuroscience
One of the most extensively studied members of the Ras superfamily of small GTPases, Rac1 is an intracellular signal transducer that remodels actin and phosphorylation signaling networks. Previous studies have shown that Rac1-mediated signaling is associated with hippocampal-dependent working memory and longer-term forms of learning and memory and that Rac1 can modulate forms of both pre- and postsynaptic plasticity. How these different cognitive functions and forms of plasticity mediated by Rac1 are linked, however, is unclear. Here, we show that spatial working memory in mice is selectively impaired following the expression of a genetically encoded Rac1 inhibitor at presynaptic terminals, while longer-term cognitive processes are affected by Rac1 inhibition at postsynaptic sites. To investigate the regulatory mechanisms of this presynaptic process, we leveraged new advances in mass spectrometry to identify the proteomic and post-translational landscape of presynaptic Rac1 signaling. We identified serine/threonine kinases and phosphorylated cytoskeletal signaling and synaptic vesicle proteins enriched with active Rac1. The phosphorylated sites in these proteins are at positions likely to have regulatory effects on synaptic vesicles. Consistent with this, we also report changes in the distribution and morphology of synaptic vesicles and in postsynaptic ultrastructure following presynaptic Rac1 inhibition. Overall, this study reveals a previously unrecognized presynaptic role of Rac1 signaling in cognitive processes and provides insights into its potential regulatory mechanisms.
