Proteomic Approach to Identify Phosphorylation-Dependent Targets of BRCT Domains; Annual rept. 1 Mar 2007-28 Feb 2008

摘要

BRCA1 C-terminal (BRCT) domains are novel phosphopeptide binding modules. Cancer-associated missense and deletion mutations have been found in the BRCT repeat regions of BRCA1, suggesting an essential role of BRCT domains in regulating BRCA1activity. In addition, BRCT domains are found in many proteins that regulate DNA damage repair, cell cycle, and genome stability, implying a more global role of BRCT domains in genome stability surveillance. These results suggest that the BRCT domain acts as a sensor to protein phosphorylation in response to DNA damage, recruits phosphorylated cellular targets, and mediates signaling complex formation. However, the identities of the in vivo BRCT domain targets are largely unknown. In order to understand the role of phosphorylation in protein-protein interactions, we developed several approaches utilizing peptide libraries and peptide arrays. We propose to use these methods to systematically identify phosphorproteins that can interact with BRCT domains. In addition to potential new regulators of genome stability, the approaches can identify phosphorylated sequences on proteins that are important for DNA damage responses and cell cycle. Such information should prove valuable, especially for the development of new screening strategies, drug targets, and treatment for breast cancer.