Mutation of Methionine 178 to Isoleucine of HIV-1 Integrase for analyzing its interaction with LEDGF/p75

Shazia, Siddiqui (2014) Mutation of Methionine 178 to Isoleucine of HIV-1 Integrase for analyzing its interaction with LEDGF/p75. Masters thesis, Indian Institute of Technology.

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Abstract

HIV-1 replication depends on the insertion of a cDNA copy of the viral genome into the host genome. This process is catalyzed by the viral integrase (IN), in a two-step process- 3’processing and strand transfer reaction. Lens epithelium-derived growth factor (LEDGF)/p75 is an important cellular co-factor for human immunodeficiency virus-1 (HIV- 1) replication which plays a crucial role during HIV-1 cDNA integration. The C-terminal integrase binding domain (IBD) of LEDGF/p75 interacts with CCD of HIV-1 Integrase (IN). In this study, based on the co-crystal structure of IN-LEDGF/p75, Methionine residue at position 178 of HIV-1 IN has been mutated to isoleucine. The M178I mutant IN awas analyzed for interaction with IBD of LEDGF/p75 in the pull down assays. Compared to wild type IN, M178I mutant show a ~90% reduction in interaction with IBD. Pull down result shows that M178I is an important residue which participates in interaction with IBD of LEDGF/p75.

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