title: [Mutations in various functional domains of HPV2 E2 protein inhibit the transcriptional depression activities].
abstract: OBJECTIVE
  To study the potential transcriptional 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 transcriptional 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 transcriptional 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 transcriptional depression function.
  
  
  CONCLUSIONS
  The transcriptional 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.
authors: State key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
authors: Chen
authors: C
authors: Gao
authors: Yan-Jun
authors: YJ
authors: Lei
authors: Hui-Ying
authors: HY
authors: Jiang
authors: Qi
authors: Q
authors: Shi
authors: Chan
authors: C
authors: Tian
authors: Jun
authors: J
authors: Han
authors: Xiao-Ping
authors: XP
authors: Dong
mesh_terms: D006367:HeLa Cells; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics*
chemical_list: D009856:Oncogene Proteins, Viral