In GFP with high risk HPV-18E6 fusion protein expressed 293T and MCF-7 cells, the endogenous wild-type p53 could be transiently phosphorylated at multiple sites

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• 作者：Lina Sun (1) (2)
  Ge Zhang (3) (4)
  Zongfang Li (3)
  Tusheng Song (5)
  Chen Huang (5)
  Lusheng Si (1) (6)
  
• 刊名：Journal of Experimental & Clinical Cancer Research
• 出版年：2008
• 出版时间：December 2008
• 年：2008
• 卷：27
• 期：1
• 全文大小：882KB
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• 作者单位：Lina Sun (1) (2)
  Ge Zhang (3) (4)
  Zongfang Li (3)
  Tusheng Song (5)
  Chen Huang (5)
  Lusheng Si (1) (6)
  
  1. Key Laboratory of Environment and Genes Related to Diseases of the Education Ministry, School of Medicine,Xi an Jiaotong University, Xi an, PR China
  2. State key Laboratory for Infectious Disease Prevention and Control, National Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, PR China
  3. The second affiliated hospital, Xi an Jiaotong University, Xi an, PR China
  4. The first hospital of Shanxi Medical University, Taiyuan, PR China
  5. Key Laboratory of Environment and Genes Related to Diseases of the Education Ministry/Department of Genetics and Molecule Biology, School of Medicine,Xi an Jiaotong University, Xi an, PR China
  6. School of Life Science and Technology ,Xi an Jiaotong University, Xi an, PR China
  
• ISSN：1756-9966

文摘

Background Infected cells recognize viral replication as a DNA damage stress and elicit the host surveillance mechanism to anti-virus infection. Modulation of the activity of tumor suppressor p53 is a key event in the replication of many viruses. They could manipulate p53 function through phosphorylation modification for their own purpose. But there is rarely research about p53 phosphorylation status in the context of HPV-E6. Therefore, we investigated whether p53 could be phosphorylated by HPV-E6. Methods We used a mammalian green fluorescence protein (GFP) expression system to express HPV-18E6 with GFP fusion proteins (GFP-18E6) in wild-type (wt) p53 cell lines, such as 293T and MCF-7 cells to trace the traffic and subcellular location of E6 protein. By immunofluorescence technique and immunoblotting, we determined the positive phosphorylated sites of p53 and observed the distribution of phosphorylated p53 in the context of GFP-18E6. Results GFP-18E6 was predominantly located in nuclei of wt p53 cell lines, and it could induce transient phosphorylation of p53 at multiple sites, such as Ser15, Ser20, and Ser392. All the three sites of phosphorylated p53s were localized in nuclei together with GFP-18E6. Conclusion In GFP with high risk HPV-18E6 fusion protein expressed 293T and MCF-7 cells, the endogenous wt p53 could be transiently phosphorylated at multiple sites.