Involvement of BRCA2 Repeats in RAD51 Mediated DNA Repair
Abstract
The overall goal of my grant proposal is to study how BRCA2 is involved in repairing DNA damage. A better understanding of BRCA2's role in breast cancer could prevent cancer formation and reduce resistance to cancer therapies. To test our hypothesis, we proposed to accomplish the following specific aims. Aim 1. To test the importance of BRC repeats in BRCA2 for binding to Rad5l in response to DNA damage. Aim2. To determine the critical residues in the BRC repeats of BRCA2 and the significance of these residues for BRCA2/Rad51 1 interactions. We screened for BRC repeat mutant that fail to bind to RAD51 1 through random mutagenesis, and one of the mutants competitively inhibited BRC-RAD5l binding was specifically identified and used for sebsequent analysis. We established cell lines tetracycline-controlled inducibly expressing of wild -type BRC repeat-UFP fusion peptides or mutant BRC repeat-GFP fusion peptide in MCF7 cells. We found that endogenous BRCA2-RAD5l binding was abolished by wild-type BRC repeat but not mutant BRC repeats. The characterization of BRC repeat inducible expression cell lines in DNA damage response and DNA repair will represent an important step toward our goal in understanding the significance of the interaction between BRCA2 and RAD5l repair complex.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 01, 2001
- Accession Number
- ADA400499
Entities
People
- Qing Zhong
Organizations
- University of Texas Health Science Center at San Antonio