Biological impacts of“hot-spot” mutations of hepatitis B virus X proteins are genotype B and C differentiated

AIM： To investigate the biological impacts of “hot-spot” mutations on genotype B and C HBV X proteins （HBx）. METHODS： Five types of “hot-spot” mutations of genotype B or C HBV X genes, which sequentially lead to the amino acid substitutions of HBx as I127T, F132Y, K130M＋V131I, I127T＋K130M＋V131I, or...

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Published in:World journal of gastroenterology : WJG 2005-08, Vol.11 (30), p.4703-4708
Main Authors: Lin, Xu, Xu, Xiao, Huang, Qing-Ling, Liu, Yu-Qing, Zheng, Da-Li, Chen, Wan-Nan, Lin, Jian-Yin
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
B基因型
Cell Division
Cell Line
C基因型
DNA, Viral - genetics
Genes, Viral
Genotype
Hepatitis B virus - genetics
Hepatitis B virus - pathogenicity
Humans
Molecular Sequence Data
Mutagenesis, Site-Directed
Mutation
Point Mutation
Recombinant Proteins - genetics
Trans-Activators - genetics
Transcriptional Activation
Transfection
Viral Hepatitis
X蛋白质
乙型肝炎病毒
基因突变
生物学研究
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Summary:AIM： To investigate the biological impacts of “hot-spot” mutations on genotype B and C HBV X proteins （HBx）. METHODS： Five types of “hot-spot” mutations of genotype B or C HBV X genes, which sequentially lead to the amino acid substitutions of HBx as I127T, F132Y, K130M＋V131I, I127T＋K130M＋V131I, or K130M＋V131I＋F132Y, respectively, were generated by means of site-directed mutagenesis. To evaluate the anti-proliferative effects, HBx or related mutants＇ expression vectors were transfected separately to the Chang cells by lipofectamine, and the cells were cultured in hygromycin selective medium for 14 d, drugresistant colonies were fixed with cold methanol, stained with Giemsa dyes and scored （increase of the colonies indicated the reduction of the anti-proliferation activity, and vice versa）. Different types of HBx expression vectors were co-transfected separately with the reporter plasmid pCMVβ to Chang cells, which were lysed 48 h posttransfection and the intra-cellular β-galactosidase activities were monitored （increase of the β-galactosidase activities indicated the reduction of the transactivation activity, and vice versa）. All data obtained were calculated by paired-samples t-test. RESULTS： As compared to standard genotype B HBx, mutants of I127T and I127T＋K130M＋V131I showed higher transactivation and anti-proliferative activities, while the mutants of F132Y, K130M＋V131I, and K130M＋V131I＋F132Y showed lower activities. As compared to standard genotype C HBx, I127T mutant showed higher transactivation activity, while the other four types of mutants showed no differences. With regard to anti-proliferative activity, compared to standard genotype C HBx, F132Y and K130M＋V131I mutants showed lower activities, and K130M＋V131I ＋F132Y mutant, on the other hand, showed higher activity, while the mutants of I127T and I127T＋K130M＋V131I showed no differences. CONCLUSION： “Hot-spot” mutations affect the antiproliferation and transactivation activities of genotype B and/or C HBx, and the biological impacts of most “hot-spot” mutations on HBx are genotype B and C differentiated.
ISSN:1007-9327
2219-2840
DOI:10.3748/wjg.v11.i30.4703