Grants and Contracts Details
Poly(ADP-ribose) polymerase (PARP-I) is a natural DNA damage biosensor whose activity triggers cellular responses leading to DNA repair or cell death by apoptosis or necrosis. Knockout experiments in mice have provided compelling evidence for PARP-I as a target for the development of radio- and chemosensitizing agents for cancer therapy including lung cancer. In the past two years, it has become apparent that PARP-I is but one of a family of enzymes with PARP activity, all containing a conserved NAD+ binding site and known inhibitors of PARP-l inhibit these enzymes. Thus, while PARP-l remains a promising therapeutic target, the discovery of multiple PARPs raises questions of inhibitor selectivity. We propose a paradigm shift in PARP-l inhibitor discovery by targeting a its unique mechanism of activation for the design of selective inhibitors. The hypothesis to be tested is that PARP-l activation. resulting from DNA strand break recognition. is mediated by a Srchomology 3 module (SH3). We will focus on the identification of the protein interactions involved in PARP-I activation and in particular whether this interaction involves an SH3 module. Specific Aim 1: To utilize co-expression of GST/His tagged PARP-l domains along with pull down assays and surface plasmon resonance techniques to identify protein interactions between PARP-I domains necessary for PARP-l activation. Specific Aim 2: We will use a structure-based approach to screen for point mutations that generate activation deficient PARP-l. These mutations are designed based on the know structure of a typical SH3 domain and are predicted to disrupt the interaction between PARP-I-SH3 module and/or the PARP-I-SH3 ligand domains.
|Effective start/end date||10/1/01 → 9/30/04|
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