Investigation og the effects of HIV-1 regulatory genes on African Green Monkey cell lines master of science

Abstract

The inheritance of retroviral genome by their hosts first gained its importance by the discovery of endogenous retroviruses in late 1960s. The subject has moved forward with the discovery of gag-like Fv1 protein which is an endogenous factor in mice that restricts MLV infection. In view of the fact that endogenous retroviruses comprise 8-10 % of primate genomes, it is well to consider that cellular expression of retroviral restriction factors may depend on retroviral regulator genes. The products of two such genes in HIV-1, tat and rev, are absolutely required for the replication of HIV-1. It was aimed to investigate the effects of HIV-1 Tat and Rev proteins on the gene expression of African Green Monkey (AGM) cells which are known to be resistant to HIV-1 infection, in an attempt to identify possible HIV restriction factors encoded by endogenous retroviral sequences.To this end, we generated stable cell lines of AGM expressing HIV-1 regulatory genes tat and rev. The expressions were confirmed by both RT-PCR at mRNA level and Western Blotting at protein level using polyclonal antibodies raised in rabbits by immunization with GST fusion proteins of Tat and Rev. Currently, we are using these cell lines for the identification of possible AGM genes selectively expressed in the presence of HIV-1 Tat and Rev proteins by microarray analysis and 2D-PAGE.