Analysis of Antiviral Roles of p53 in the Early Replication of Retrovirus
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Issue Date
2014
Authors
Alghamdi, Faris Saeed '14
Degree
MS in Biotechnology
Advisor
Shi, Binshan
Committee Members
Yager, Eric
Stein, Markus
Stein, Markus
Journal Title
Journal ISSN
Volume Title
Abstract
The tumor suppressor protein p53 has been known as "the guardian of the genome". p53 functions to maintain the host cell genome integrity by arresting cell cycle, activating DNA repair, and inducing apoptosis. Research has demonstrated that retroviruses interact with various host cell proteins involved in the cell cycle regulation and DNA repair machinery. However, the possible influence of p53 in the replication of retroviruses has not been well studied. In order to investigate the role of p53 in retrovirus early replication, Murine leukemia virus (MLV) vector based retroviruses were produced and used to infect human HCT 116 p53+/+ cells and HCT 116 p53-/- cells. MLV was produced by co-transfection of GP2 293 packaging cells by pRetroZsGreen1 and pVSV-G plasmids. Real time PCR quantification showed that the produced retroviruses titer was up to 3.23x108 copies/ml. The infection of MLV was performed in both dividing HCT116 cells and non-dividing HCT116 cells cultured by prolonged serum depletion. The result showed that there were no obvious difference in infection rate, amount of late reverse transcription (RT) product, and circular viral DNA between the dividing HCT116 p53+/+ and dividing HCT116 p53 -/- cells; indicating that p53 does not block the replication of MLV in dividing cells. However, it was found that the infection rate in non-dividing HCT 116 p53-/- (56%) doubled the infection rate in non-dividing HCT116 p53+/+cells (28%). Meanwhile, the amounts of late RT, circular viral 1-LTR, and 2-LTR DNA were significantly higher in non-dividing HCT116 p53-/- compared with non-dividing HCT 116 p53+/+ cells. This result strongly suggests that p53 inhibits MLV early replication in non-dividing cells. It was also evidenced that p53 promotes the formation of viral 1 LTR circular DNA in both dividing and no-dividing cells. Our data implies p53 might be an important host cellular factor in defending retrovirus infection in non-dividing cells.
Citation
Alghamdi, Faris. "Analysis of Antiviral Roles of P53 in the Early Replication of Retrovirus." Albany College of Pharmacy and Health Sciences, New York, Proquest/UMI, 2014.
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