HPV-16E6 can induce multiple site phosphorylation of p53.
Modulation of the activity of tumor suppressor p53 is a key event in the replication of many viruses. They could manipulate p53 function through modification of phosphorylation for their own purpose. However, there are scarce data on the relationship between high risk human papillomavirus (HPV) E6 protein and p53 phosphorylation status. Therefore, we used a mammalian green fluorescence protein (GFP) expression system to express HPV-16E6 with GFP fusion proteins in wild-type p53 cell lines, 293T, MCF-7, and SMMC-7721 to trace the traffic and subcellular location of E6 protein. By immunoblotting, we determined the positive phosphorylated sites of p53 in the context of HPV-16E6. Using immunofluorescence techniques, we observed the distribution of phosphorylated p53 in all the cells we used. In conclusion, HPV-16E6 was predominantly located in nuclei of wild-type p53 cells, and it was able to induce phosphorylation of p53 at multiple sites, such as Ser15, Ser20, and Ser392. The level and time of these phosphorylated sites of p53 were different in HPV-16E6 expressing cells. Furthermore, the phosphorylated p53 was localized in the nuclei together with HPV-16E6.
Zhang
Ge
G
The Secondary Affiliated Hospital of School of Medicine, Xi'an, People's Republic of China.
Sun
Lina
L
Li
Zongfang
Z
Si
Lusheng
L
Song
Tusheng
T
Huang
Chen
C
Zhang
Wanggang
W
D015153:Blotting, Western; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000821:virology; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins; D052162:Human papillomavirus 16 / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins / Q000378:metabolism*; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins