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Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane.

Publication ,  Journal Article
Hannigan, MM; Hoffman, AM; Thompson, JW; Zheng, T; Nicchitta, CV
Published in: Mol Cell Proteomics
November 2020

Protein synthesis on the endoplasmic reticulum (ER) requires the dynamic coordination of numerous cellular components. Together, resident ER membrane proteins, cytoplasmic translation factors, and both integral membrane and cytosolic RNA-binding proteins operate in concert with membrane-associated ribosomes to facilitate ER-localized translation. Little is known, however, regarding the spatial organization of ER-localized translation. This question is of growing significance as it is now known that ER-bound ribosomes contribute to secretory, integral membrane, and cytosolic protein synthesis alike. To explore this question, we utilized quantitative proximity proteomics to identify neighboring protein networks for the candidate ribosome interactors SEC61β (subunit of the protein translocase), RPN1 (oligosaccharyltransferase subunit), SEC62 (translocation integral membrane protein), and LRRC59 (ribosome binding integral membrane protein). Biotin labeling time course studies of the four BioID reporters revealed distinct labeling patterns that intensified but only modestly diversified as a function of labeling time, suggesting that the ER membrane is organized into discrete protein interaction domains. Whereas SEC61β and RPN1 reporters identified translocon-associated networks, SEC62 and LRRC59 reporters revealed divergent protein interactomes. Notably, the SEC62 interactome is enriched in redox-linked proteins and ER luminal chaperones, with the latter likely representing proximity to an ER luminal chaperone reflux pathway. In contrast, the LRRC59 interactome is highly enriched in SRP pathway components, translation factors, and ER-localized RNA-binding proteins, uncovering a functional link between LRRC59 and mRNA translation regulation. Importantly, analysis of the LRRC59 interactome by native immunoprecipitation identified similar protein and functional enrichments. Moreover, [35S]-methionine incorporation assays revealed that siRNA silencing of LRRC59 expression reduced steady state translation levels on the ER by ca. 50%, and also impacted steady state translation levels in the cytosol compartment. Collectively, these data reveal a functional domain organization for the ER and identify a key role for LRRC59 in the organization and regulation of local translation.

Duke Scholars

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Published In

Mol Cell Proteomics

DOI

EISSN

1535-9484

Publication Date

November 2020

Volume

19

Issue

11

Start / End Page

1826 / 1849

Location

United States

Related Subject Headings

  • SEC Translocation Channels
  • Ribosomes
  • Recombinant Proteins
  • RNA, Small Interfering
  • Proteomics
  • Proteome
  • Protein Interaction Maps
  • Protein Biosynthesis
  • Proteasome Endopeptidase Complex
  • Oxidation-Reduction
 

Citation

APA
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Hannigan, M. M., Hoffman, A. M., Thompson, J. W., Zheng, T., & Nicchitta, C. V. (2020). Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane. Mol Cell Proteomics, 19(11), 1826–1849. https://doi.org/10.1074/mcp.RA120.002228
Hannigan, Molly M., Alyson M. Hoffman, J Will Thompson, Tianli Zheng, and Christopher V. Nicchitta. “Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane.Mol Cell Proteomics 19, no. 11 (November 2020): 1826–49. https://doi.org/10.1074/mcp.RA120.002228.
Hannigan MM, Hoffman AM, Thompson JW, Zheng T, Nicchitta CV. Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane. Mol Cell Proteomics. 2020 Nov;19(11):1826–49.
Hannigan, Molly M., et al. “Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane.Mol Cell Proteomics, vol. 19, no. 11, Nov. 2020, pp. 1826–49. Pubmed, doi:10.1074/mcp.RA120.002228.
Hannigan MM, Hoffman AM, Thompson JW, Zheng T, Nicchitta CV. Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane. Mol Cell Proteomics. 2020 Nov;19(11):1826–1849.

Published In

Mol Cell Proteomics

DOI

EISSN

1535-9484

Publication Date

November 2020

Volume

19

Issue

11

Start / End Page

1826 / 1849

Location

United States

Related Subject Headings

  • SEC Translocation Channels
  • Ribosomes
  • Recombinant Proteins
  • RNA, Small Interfering
  • Proteomics
  • Proteome
  • Protein Interaction Maps
  • Protein Biosynthesis
  • Proteasome Endopeptidase Complex
  • Oxidation-Reduction