Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

Xiaojun Wu; Jennifer S. Spence; Tandrila Das; Xiaoqiu Yuan; Chengjie Chen; Yuqing Zhang; Yumeng Li; Yanan Sun; Kartik Chandran; Howard C. Hang; Tao Peng

doi:10.1016/j.chembiol.2020.03.004

Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

Xiaojun Wu, Jennifer S. Spence, Tandrila Das, Xiaoqiu Yuan, Chengjie Chen, Yuqing Zhang, Yumeng Li, Yanan Sun, Kartik Chandran, Howard C. Hang, Tao Peng

Microbiology & Immunology

Research output: Contribution to journal › Article

Abstract

Interferon-induced transmembrane protein 3 (IFITM3) is a key interferon effector that broadly prevents infection by diverse viruses. However, the cellular factors that control IFITM3 homeostasis and antiviral activity have not been fully elucidated. Using site-specific photo-crosslinking and quantitative proteomic analysis, here we present the identification and functional characterization of VCP/p97 AAA-ATPase as a primary interaction partner of IFITM3. We show that IFITM3 ubiquitination at lysine 24 is crucial for VCP binding, trafficking, turnover, and engagement with incoming virus particles. Consistently, pharmacological inhibition of VCP/p97 ATPase activity leads to defective IFITM3 lysosomal sorting, turnover, and co-trafficking with virus particles. Our results showcase the utility of site-specific protein photo-crosslinking in mammalian cells and reveal VCP/p97 as a key cellular factor involved in IFITM3 trafficking and homeostasis.

Original language	English (US)
Pages (from-to)	571-585.e6
Journal	Cell Chemical Biology
Volume	27
Issue number	5
DOIs	https://doi.org/10.1016/j.chembiol.2020.03.004
State	Published - May 21 2020

Keywords

IFITM3
VCP/p97
chemical proteomics
photo-crosslinking
protein-protein interaction
unnatural amino acid

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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Wu, X., Spence, J. S., Das, T., Yuan, X., Chen, C., Zhang, Y., Li, Y., Sun, Y., Chandran, K., Hang, H. C., & Peng, T. (2020). Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase. Cell Chemical Biology, 27(5), 571-585.e6. https://doi.org/10.1016/j.chembiol.2020.03.004

Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase. / Wu, Xiaojun; Spence, Jennifer S.; Das, Tandrila; Yuan, Xiaoqiu; Chen, Chengjie; Zhang, Yuqing; Li, Yumeng; Sun, Yanan; Chandran, Kartik; Hang, Howard C.; Peng, Tao.

In: Cell Chemical Biology, Vol. 27, No. 5, 21.05.2020, p. 571-585.e6.

Research output: Contribution to journal › Article

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abstract = "Interferon-induced transmembrane protein 3 (IFITM3) is a key interferon effector that broadly prevents infection by diverse viruses. However, the cellular factors that control IFITM3 homeostasis and antiviral activity have not been fully elucidated. Using site-specific photo-crosslinking and quantitative proteomic analysis, here we present the identification and functional characterization of VCP/p97 AAA-ATPase as a primary interaction partner of IFITM3. We show that IFITM3 ubiquitination at lysine 24 is crucial for VCP binding, trafficking, turnover, and engagement with incoming virus particles. Consistently, pharmacological inhibition of VCP/p97 ATPase activity leads to defective IFITM3 lysosomal sorting, turnover, and co-trafficking with virus particles. Our results showcase the utility of site-specific protein photo-crosslinking in mammalian cells and reveal VCP/p97 as a key cellular factor involved in IFITM3 trafficking and homeostasis.",

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