Mutations in various functional domains of HPV2 E2 protein inhibit the transcriptional depression activities
10.3760/cma.j.issn.1003-9279.2011.03.002
- VernacularTitle:人乳头瘤病毒2(HPV2)E2蛋白不同功能区突变对转录抑制作用影响的研究
- Author:
Chen GAO
1
;
Yan-Jun LEI
;
Hui-Ying JIANG
;
Qi SHI
;
Chan TIAN
;
Jun HAN
;
Xiao-Ping DONG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Papillomavirus,human;
Viralproteins;
Mutation;
Promoter regions (genetics)
- From:
Chinese Journal of Experimental and Clinical Virology
2011;25(3):164-166
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the potential transcriptionai depression activities of HPV2 E2 proteins with mutations in different functional domains. Methods The primers for constructing various E2 mutants were synthesized based on a HPV2 isolate containing several point mutations within E2 open reading frame. Different E2 mutations were generated by the method of extending PCR and inserted into plasmid pcDNA3. 1. Various recombinant mammalian expression plasmids pcDNA3. 1-E2 were co-transfected into HeLa cells together with a CAT-reporter plasmid pBLCAT-LCR containing HPV-2 prototype LCR, respectively. The transcriptionai repression activities of the E2 mutants were evaluated by detection of CAT expression values. Results Compared with the full-length prototype E2, removals of both N- and C-terminal domains abolished E2 transcriptionai repressive activities. The point mutations in the transactivation domain ( nt 3037 ) , the internal hinge region ( nt 3387 ) and DNA binding domain ( nt 3697 ) showed remarkable inhibition on its transcriptionai depression function. Conclusion The transcriptionai regulation activity of HPV2 E2 is related with its DNA binding and transactivation domains. The exchanges of the single amino acid within E2, derived from a HPV2 isolate, abolish significantly the repressive effect on viral promoter in the context of full-length E2.
