Proximity proteomics identifies novel function of Rab14 in trafficking of Ebola virus matrix protein VP40

Jiannan Fan; Xuechun Liu; Feifei Mao; Xihua Yue; Intaek Lee; Yingke Xu

doi:10.1016/j.bbrc.2020.04.041

Proximity proteomics identifies novel function of Rab14 in trafficking of Ebola virus matrix protein VP40

Biochem Biophys Res Commun. 2020 Jun 25;527(2):387-392. doi: 10.1016/j.bbrc.2020.04.041. Epub 2020 Apr 21.

Authors

Jiannan Fan¹, Xuechun Liu¹, Feifei Mao², Xihua Yue², Intaek Lee², Yingke Xu³

Affiliations

¹ Department of Biomedical Engineering, Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, 310027, China.
² School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China.
³ Department of Biomedical Engineering, Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, 310027, China; School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China; Department of Endocrinology, The Affiliated Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310016, China. Electronic address: yingkexu@zju.edu.cn.

PMID: 32327259
DOI: 10.1016/j.bbrc.2020.04.041

Abstract

Ebola virus is a member of Filoviridae family of viruses that causes fetal hemorrhagic fever in human. Matrix protein VP40 of the Ebola virus is involved in multiple stages of viral maturation processes. In order to fully understand the interacting partners of VP40 in host cells, we applied proximity-dependent biotin-identification (BioID) approach to systematically screen for potential proteins at different time points of VP40 expression. By immunoprecipitation and subsequent proteomics analysis, we found over 100 candidate proteins with various cellular components and molecular functions. Among them, we identified Rab14 GTPase that appears to function at the late stage of VP40 expression. Imaging studies demonstrated that VP40 and Rab14 have substantial colocalization when expressed in HeLa cells. Overexpression of the dominant-negative Rab14(S25N) diminished the plasma membrane (PM) localization of VP40. In addition, we found that secreted VP40 protein can be endocytosed into Rab14 positive compartments. In summary, our study provides evidence that Rab14 is a novel regulator of the intracellular trafficking of Ebola virus matrix protein VP40 in HeLa cells.

Keywords: BioID; Ebola virus; Proteomics; Rab14; VP40.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ebolavirus / physiology*
HeLa Cells
Hemorrhagic Fever, Ebola / metabolism*
Host-Pathogen Interactions*
Humans
Nucleoproteins / metabolism*
Protein Interaction Maps
Protein Transport
Viral Core Proteins / metabolism*
rab GTP-Binding Proteins / metabolism*

Substances

Nucleoproteins
Viral Core Proteins
nucleoprotein VP40, Ebola virus
Rab14 protein, human
rab GTP-Binding Proteins