TY - JOUR
T1 - USP7 substrates identified by proteomics analysis reveal the specificity of USP7
AU - Nie, Litong
AU - Wang, Chao
AU - Liu, Xiaoguang
AU - Teng, Hongqi
AU - Li, Siting
AU - Huang, Min
AU - Feng, Xu
AU - Pei, Guangsheng
AU - Hang, Qinglei
AU - Zhao, Zhongming
AU - Gan, Boyi
AU - Ma, Li
AU - Chen, Junjie
N1 - Publisher Copyright:
© 2022 Nie et al.
PY - 2022/9/1
Y1 - 2022/9/1
N2 - Deubiquitylating enzymes (DUBs) remove ubiquitin chains from proteins and regulate protein stability and function. USP7 is one of the most extensively studied DUBs, since USP7 has several well-known substrates important for cancer progression, such as MDM2, N-MYC, and PTEN. Thus, USP7 is a promising drug target. However, systematic identification of USP7 substrates has not yet been performed. In this study, we carried out proteome profiling with label-free quantification in control and single/double-KO cells of USP7 and its closest homolog, USP47. Our proteome profiling for the first time revealed the proteome changes caused by USP7 and/or USP47 depletion. Combining protein profiling, transcriptome analysis, and tandem affinity purification of USP7-associated proteins, we compiled a list of 20 high-confidence USP7 substrates that includes known and novel USP7 substrates. We experimentally validated MGA and PHIP as new substrates of USP7. We further showed that MGA deletion reduced cell proliferation, similar to what was observed in cells with USP7 deletion. In conclusion, our proteome-wide analysis uncovered potential USP7 substrates, providing a resource for further functional studies.
AB - Deubiquitylating enzymes (DUBs) remove ubiquitin chains from proteins and regulate protein stability and function. USP7 is one of the most extensively studied DUBs, since USP7 has several well-known substrates important for cancer progression, such as MDM2, N-MYC, and PTEN. Thus, USP7 is a promising drug target. However, systematic identification of USP7 substrates has not yet been performed. In this study, we carried out proteome profiling with label-free quantification in control and single/double-KO cells of USP7 and its closest homolog, USP47. Our proteome profiling for the first time revealed the proteome changes caused by USP7 and/or USP47 depletion. Combining protein profiling, transcriptome analysis, and tandem affinity purification of USP7-associated proteins, we compiled a list of 20 high-confidence USP7 substrates that includes known and novel USP7 substrates. We experimentally validated MGA and PHIP as new substrates of USP7. We further showed that MGA deletion reduced cell proliferation, similar to what was observed in cells with USP7 deletion. In conclusion, our proteome-wide analysis uncovered potential USP7 substrates, providing a resource for further functional studies.
KW - MGA
KW - proteomics
KW - ubiquitination
KW - USP7
UR - http://www.scopus.com/inward/record.url?scp=85141894070&partnerID=8YFLogxK
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U2 - 10.1101/gad.349848.122
DO - 10.1101/gad.349848.122
M3 - Article
C2 - 36302555
AN - SCOPUS:85141894070
SN - 0890-9369
VL - 36
SP - 1016
EP - 1030
JO - Genes and Development
JF - Genes and Development
IS - 17-18
ER -