Quantitative proteomics reveal proteins enriched in tubular endoplasmic reticulum of Saccharomyces cerevisiae

  1. Xinbo Wang
  2. Shanshan Li
  3. Haicheng Wang
  4. Wenqing Shui  Is a corresponding author
  5. Junjie Hu  Is a corresponding author
  1. Nankai University, China
  2. Shanghai Tech University, China
  3. Huazhong University of Science and Technology, China
  4. Chinese Academy of Sciences, China

Abstract

The tubular network is a critical part of the endoplasmic reticulum (ER). It is shaped by the reticulons and REEPs/Yop1p that generate tubules by inducing high membrane curvature, and the dynamin-like GTPases atlastin and Sey1p/RHD3 that connect tubules via membrane fusion. However, the specific functions of this ER domain are not clear. Here, we isolated tubule-based microsomes from Saccharomyces cerevisiae via classical cell fractionation and detergent-free immunoprecipitation of Flag-tagged Yop1p, which specifically localizes to ER tubules. In quantitative comparisons of tubule-derived and total microsomes, we identified a total of 79 proteins that were enriched in the ER tubules, including known proteins that organize the tubular ER network. Functional categorization of the list of proteins revealed that the tubular ER network may be involved in membrane trafficking, lipid metabolism, organelle contact, and stress sensing. We propose that affinity isolation coupled with quantitative proteomics is a useful tool for investigating ER functions.

Article and author information

Author details

  1. Xinbo Wang

    Department of Genetics and Cell Biology, Nankai University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Shanshan Li

    iHuman Institute, Shanghai Tech University, Shanghai, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Haicheng Wang

    College of Life Sciences, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Wenqing Shui

    iHuman Institute, Shanghai Tech University, Shanghai, China
    For correspondence
    shuiwq_tib@163.com
    Competing interests
    The authors declare that no competing interests exist.
  5. Junjie Hu

    National Laboratory of Biomacromolecules, Chinese Academy of Sciences, Beijing, China
    For correspondence
    huj@ibp.ac.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4712-2243

Funding

Howard Hughes Medical Institute (International Early Career Scientist grant)

  • Junjie Hu

National Natural Science Foundation of China (31630020)

  • Junjie Hu

Ministry of Science and Technology of the People's Republic of China (2016YFA0500201)

  • Junjie Hu

National Natural Science Foundation of China (31401150)

  • Wenqing Shui

Chinese Academy of Sciences (Bairenjihua Program)

  • Wenqing Shui

National Natural Science Foundation of China (3142100024)

  • Junjie Hu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Randy Schekman, Howard Hughes Medical Institute, University of California, Berkeley, United States

Publication history

  1. Received: December 1, 2016
  2. Accepted: March 6, 2017
  3. Accepted Manuscript published: March 13, 2017 (version 1)
  4. Version of Record published: March 20, 2017 (version 2)

Copyright

© 2017, Wang 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.

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  1. Xinbo Wang
  2. Shanshan Li
  3. Haicheng Wang
  4. Wenqing Shui
  5. Junjie Hu
(2017)
Quantitative proteomics reveal proteins enriched in tubular endoplasmic reticulum of Saccharomyces cerevisiae
eLife 6:e23816.
https://doi.org/10.7554/eLife.23816
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