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

Abstract
Article and author information
Metrics

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

Xinbo Wang

Department of Genetics and Cell Biology, Nankai University, Tianjin, China

Competing interests
The authors declare that no competing interests exist.
Shanshan Li

iHuman Institute, Shanghai Tech University, Shanghai, China

Competing interests
The authors declare that no competing interests exist.
Haicheng Wang

College of Life Sciences, Huazhong University of Science and Technology, Wuhan, China

Competing interests
The authors declare that no competing interests exist.
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.
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.

"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.

Copyright

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.