Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics

doi:10.1021/acs.jproteome.9b00139

	Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics
	Chen, Xiangling 1,2,3; Lu, Dayun 1,2,3; Gao, Jing 1,2; Zhu, Hongwen 1,2; Zhou, Yanting 1,2; Gao, Daming 3,4; Zhou, Hu 1,2,3
刊名	JOURNAL OF PROTEOME RESEARCH
	2019-06-01
卷号	18 期号:6 页码:2654-2665
关键词	deubiquitinase USP9X NFX1-123 SILAC quantitative proteomics
ISSN号	1535-3893
DOI	10.1021/acs.jproteome.9b00139
通讯作者	Zhou, Hu(zhouhu@simm.ac.cn)
英文摘要	The deubiquitinase USP9X is involved in multiple diseases including neurodegeneration, epilepsy, and various types of tumors by targeting different substrates. In the present study, we aimed to explore the potential substrates of USP9X and performed SILAC-based quantitative proteomics to compare these substrates in USP9X-knockdown and wild-type HeLa cells. We consequently carried out Flag-NFX1-123 tag affinity-based mass spectrometry and confirmed that the X-box binding nuclear factor NFX1-123 interacted with USP9X. Moreover, immunoprecipitation assays verified a direct interaction between USP9X and NFX1-123. Further experiments confirmed that NFX1-123 could be modified by ubiquitination and that USP9X stabilized NFX1-123 via efficient deubiquitination of NFX1-123. Knockdown of USP9X resulted in decreased NFX1-123 protein levels compared with their unchanged corresponding mRNA levels in different cell lines. In summary, we found that NFX1-123 was a bona fide substrate of the deubiquitinase USP9X and that it could be degraded by the ubiquitin-proteasome system. The present study provided new insight into understanding the biological function of USP9X by targeting its substrate NFX1-123.
资助项目	National Key Research and Development Program from the Ministry of Science and Technology of China[2017YFC1700200] ; National Natural Science Foundation of China[31800693] ; National Natural Science Foundation of China[31570830] ; Innovation Project of Instrument and Equipment Function Development from the Bureau of Goods, Chinese Academy of Sciences[YZ201542]
WOS关键词	HUMAN-PAPILLOMAVIRUS 16E6 ; DEUBIQUITINASE USP9X ; TELOMERASE ACTIVITY ; BINDING-PROTEINS ; EXPRESSION ; INTERACTS ; HPV ; UBIQUITINATION ; STABILIZATION ; TRANSCRIPTION
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000471212200028
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/289549]
专题	中国科学院上海药物研究所
通讯作者	Zhou, Hu
作者单位	1.Chinese Acad Sci, Dept Analyt Chem, Shanghai 201203, Peoples R China 2.Chinese Acad Sci, CAS Key Lab Receptor Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 3.Univ Chinese Acad Sci, 19A Yuquan Rd, Beijing 100049, Peoples R China 4.Chinese Acad Sci, Shanghai Inst Biol Sci, CAS Key Lab Syst Biol,Innovat Ctr Cell Signaling, CAS Ctr Excellence Mol Cell Sci,Inst Biochem & Ce, Shanghai 200031, Peoples R China
推荐引用方式 GB/T 7714	Chen, Xiangling,Lu, Dayun,Gao, Jing,et al. Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics[J]. JOURNAL OF PROTEOME RESEARCH,2019,18(6):2654-2665.
APA	Chen, Xiangling.,Lu, Dayun.,Gao, Jing.,Zhu, Hongwen.,Zhou, Yanting.,...&Zhou, Hu.(2019).Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics.JOURNAL OF PROTEOME RESEARCH,18(6),2654-2665.
MLA	Chen, Xiangling,et al."Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics".JOURNAL OF PROTEOME RESEARCH 18.6(2019):2654-2665.