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 › peer-review

1 Scopus citations

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

Access to Document

10.1016/j.chembiol.2020.03.004

Fingerprint Dive into the research topics of 'Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase'. Together they form a unique fingerprint.

Cite this

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 › peer-review

@article{6b240054595741e4b84a8df5d016e772,

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

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.",

keywords = "IFITM3, VCP/p97, chemical proteomics, photo-crosslinking, protein-protein interaction, unnatural amino acid",

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

note = "Funding Information: We thank Prof. Peng R. Chen at Peking University for the pCMV-DiZPK-PylRS plasmid. We also thank the Proteomics Resource Center at The Rockefeller University for assistance in proteomic analysis. T.D. and X.Y. acknowledge the Tri-Institutional Program in Chemical Biology at The Rockefeller University. K.C. acknowledges support from NIH -NIAID R01AI134824 . H.C.H. acknowledges support from NIH -NIGMS R01M087544 . T.P. acknowledges support from National Natural Science Foundation of China ( 21778010 ), Shenzhen Science and Technology Innovation Committee ( JCYJ20170412150832022 ), and Shenzhen Peacock Plan ( KQTD2015032709315529 ). ",

year = "2020",

month = may,

day = "21",

doi = "10.1016/j.chembiol.2020.03.004",

language = "English (US)",

volume = "27",

pages = "571--585.e6",

journal = "Cell Chemical Biology",

issn = "2451-9448",

publisher = "Elsevier Inc.",

number = "5",

}

TY - JOUR

T1 - Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

AU - Wu, Xiaojun

AU - Spence, Jennifer S.

AU - Das, Tandrila

AU - Yuan, Xiaoqiu

AU - Chen, Chengjie

AU - Zhang, Yuqing

AU - Li, Yumeng

AU - Sun, Yanan

AU - Chandran, Kartik

AU - Hang, Howard C.

AU - Peng, Tao

N1 - Funding Information: We thank Prof. Peng R. Chen at Peking University for the pCMV-DiZPK-PylRS plasmid. We also thank the Proteomics Resource Center at The Rockefeller University for assistance in proteomic analysis. T.D. and X.Y. acknowledge the Tri-Institutional Program in Chemical Biology at The Rockefeller University. K.C. acknowledges support from NIH -NIAID R01AI134824 . H.C.H. acknowledges support from NIH -NIGMS R01M087544 . T.P. acknowledges support from National Natural Science Foundation of China ( 21778010 ), Shenzhen Science and Technology Innovation Committee ( JCYJ20170412150832022 ), and Shenzhen Peacock Plan ( KQTD2015032709315529 ).

PY - 2020/5/21

Y1 - 2020/5/21

N2 - 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.

AB - 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.

KW - IFITM3

KW - VCP/p97

KW - chemical proteomics

KW - photo-crosslinking

KW - protein-protein interaction

KW - unnatural amino acid

UR - http://www.scopus.com/inward/record.url?scp=85083274750&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85083274750&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2020.03.004

DO - 10.1016/j.chembiol.2020.03.004

M3 - Article

C2 - 32243810

AN - SCOPUS:85083274750

VL - 27

SP - 571-585.e6

JO - Cell Chemical Biology

JF - Cell Chemical Biology

SN - 2451-9448

IS - 5

ER -