Hepatitis B spliced protein enhanced NF-κB activities via interaction with ubiquitously expressed transcript splice variant 1
CHEN Wan-nan, YU Yan-fei, WU Qiong, LIU Shen-min, WU Long-fei, SHI Jia-jian, LIN Xu
Key Laboratory for Gastrointestinal Cancer, Ministry of Education / Key Laboratory of Fujian Province for Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, China
Abstract:We investigated whether hepatitis B spliced protein affect the NF-kappa B activities by interacting with ubiquitously expressed transcript splice variant 1 (UXT-V1). The HBSP-UXT-V1 protein interactions were screened by yeast two hybrid assay, and confirmed by immunoprecipitation, confocal microscopy, mammalian two hybrid assay, and GST-Pulldown assay. The reporter plasmids driven by NF-kappa B promoter were transfected into UXT-knockdown HBSP stably expressed cell lines, and the reporter genes were detected after transfection. Results showed that the interaction between UXT-V1 and HBSP in yeast was demonstrated. Furtherly, HBSP could interact with UXT-V1 in mammalian cells. HBSP could enhance NF-kappa B activities, and this effect was partly achieved by the interaction with UXT-V1. In conclusion, the effect of HBSP-UXT-V1 interaction on the NF-kappa B pathway in hepatocytes may have an impact on HBV related liver diseases.
[1] Lee WM. Hepatitis B virus infection[J]. N Engl J Med, 1997, 337: 1733-1745. doi:10.1056/NEJM199712113372406 [2] Rosmorduc O, Petit MA, Pol S, et al. In vivo and in vitriol expression of defective hepatitis B virus particles generated by spliced hepatitis B virus RNA[J]. Hepatology, 1995, 22(1): 10-19. doi:10.1002/hep.1840220103 [3] Mancini-Bourgine M, Bayard F, Soussan P, et al. Hepatitis B virus splice-generated protein induces T-cell responses in HLA-transgenic mice and hepatitis B virus-infected patients[J]. J Virol, 2007, 81(10): 4963-4972. doi:10.1128/JVI.02619-06 [4] Chen WN, Chen JY, Jiao BY, et al. Interaction of hepatitis B spliced protein (HBSP) with cathepsin B promotes hepatoma cell migration and invasion[J]. J Virol, 2012, 86(24): 13533-13541. doi:10.1128/JVI.02095-12 [5] Chen JY, Chen WN, Liu LL, et al. Hepatitis B spliced protein (HBSP) generated by a spliced hepatitis B virus RNA participates in abnormality of fibrin formation and functions by binding to fibrinogen γ chain[J]. J Med Virol, 2010, 82(12): 2019-2026. doi:10.1002/jmv.21918 [6] Sun S, Tang Y, Lou X, et al. UXT is a novel and essential cofactor in the NF-kappaB transcriptional enhanceosome[J]. J Cell Biol, 2007, 178(2): 231-244. doi:10.1083/jcb.200611081 [7] Huang Y, Liu H, Ge R, et al. UXT-V1 facilitates the formation of MAVS antiviral signalosome on mitochondria[J]. J Immunol, 2012, 188(1): 358-366. doi:10.4049/jimmunol.1102079 [8] Soussan P, Garreau F, Zylberberg H, et al. In vivo expression of a new hepatitis B virus protein encoded by a spliced RNA[J]. J Clin Invest, 2000,105(1): 55-60. doi:10.1172/JCI8098 [9] Soussan P, Tuveri R, Nalpas B, et al. The expression of hepatitis B spliced protein (HBSP) encoded by a spliced hepatitis B virus RNA is associated with viral replication and liver fibrosis[J]. J Hepatol, 2003, 38(3): 343-348. doi:10.1016/S0168-8278(02)00422-1 [10] Duriez M, Mandouri Y, Lekbaby B, et al. Alternative splicing of Hepatitis B virus: a novel virus/host interaction altering liver immunity[J]. J Hepatol, 2017, 67(4): 687-699. doi:10.1016/j.jhep.2017.05.025. S0168-8278(17)32065-2 [11] Chen JY, Chen WN, Jiao BY, et al. Hepatitis B Spliced Protein (HBSP) promotes the carcinogenic effects of Benzo [alpha] pyrene by interacting with microsomal epoxide hydrolase and enhancing its hydrolysis activity[J]. BMC Cancer, 2014,14(1): 282. doi:10.1186/1471-2407-14-282 [12] Chen WN, Huang JG, Liang FF, et al. Hepatitis B spliced protein triggers TGFβ1-induced epithelial-mesenchymal transition via interaction with transforming growth factor beta-1-induced transcript 1 protein[J]. Chin J Zoonoses,2017,33 (4): 305-311. doi:10.3969/j.issn.1002-2694.2017.04.003 (in Chinese) 陈婉南,黄俊高,梁菲菲,等.乙型肝炎病毒剪接特异性蛋白HBSP与TGFβ1诱导蛋白1相互作用促进TGFβ1诱导的肝癌细胞上皮间质转化[J].中国人兽共患病学报,2017 ,33 (4) :305-311. [13] Huang Y, Chen L, Zhou Y, et al. UXT-V1 protects cells against TNF-induced apoptosis through modulating complex II formation[J]. Mol Biol Cell, 2011, 22(8): 1389-1397. doi:10.1091/mbc.E10-10-0827 [14] Sethurathinam S, Singh LP, Panneerselvam P, et al. UXT plays dual opposing roles on SARM-induced apoptosis[J]. FEBS Lett,2013,587(20): 3296-3302. doi:10.1016/j.febslet.2013.08.033 [15] Taneja SS, Ha S, Swenson NK, et al. ART-27, an androgen receptor coactivator regulated in prostate development and cancer[J]. J Biol Chem,2004,279(14): 13944. doi:10.1074/jbc.M306576200 [16] Sun S. UXT is a novel and essential cofactor in the NF-κb transcriptional enhanceosome[J]. J Cell Biol, 2007, 178(2): 231-244. doi:10.1083/jcb.200611081 [17] Gilmore TD. Introduction to NF-κB: players, pathways, perspectives[J]. Oncogene, 2006, 25(51): 6680-6684. doi:10.1038/sj.onc.1209954 [18] Hoesel B,Schmid JA. The complexity of NF-κB signaling in inflammation and cancer[J]. Mol Cancer, 2013,12(1): 86. doi:10.1186/1476-4598-12-86