TcpC is not only an E3 ubiquitin ligase but also an inhibitor of USP14 deubiquitinase
作者单位:Institute of Translational MedicineHangzhou City University Department of Clinical MedicineHangzhou City University School of Medicine Department of Basic Medical SciencesZhejiang University School of Medicine
会议名称:《浙江省免疫学会第十三次学术大会》
会议日期:1000年
学科分类:1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学]
关 键 词:TcpC inhibit deubiquitinase USP14 innate immune
摘 要:Background:Toll/interleukin 1 receptor domain containing protein encoded by ***(TcpC) is a multifunctional virulence factor in the majority of uropathogenic ***(UPEC) strains and blocks the TLR signaling pathway by serving as an E3 ubiquitin ligase that promotes MyD88 degradation by ubiquitin-proteasome pathway,hereby inhibiting the TLR mediated activation of ***:This study aims to investigate if TcpC can also inhibit the activity of deubiquitinase and its related *** and methods:TcpC secreting wild-type CFT073(CFT073) was kindly provided by Professor Jian-guo *** knock out CFT073 mutant(CFT073Δtcpc) was constructed in our *** human monocytic cell line THP-1 and mouse macrophage cell lines J774 A.1 were obtained from Shanghai Institute of Biochemistry and Cell *** situ co-localization of TcpC with USP14,USP14 with ubiquitin and Rpn1 in kidneys from pyelonephritis mouse model was determined by confocal *** of UPEC in macrophages was observed by holographic 3 D live cell imaging system,flow cytometry and confocal *** of deubiquitinase and proteasome were measured by fluorescent microplate reader.K48-ubiquitination of MyD88 in rTcpC treated macrophages was detected by *** mRNA and protein expression levels of pro-inflammatory cytokines were detected by qRT-PCR and *** abilities of rTcpC with rUSP14 was determined by surface plasmon resonance(SPR) and microscale thermophoresis(MST).Transcription and protein level of ubiquitin and proteasome related genes were detected by transcriptome and proteome *** amino acids required to maintain the deubiquitinase inhibitory activity in rTcpC were identified by ***:In situ co-localization of TcpC and ubiquitin with USP14 or PSMD2 with USP14 was significantly enhanced or declined respectively in kidney from CFT073 infected mice compared to kidney isolated from mice infect