Mechanisms and Components of the DNA Damage Checkpoint
Abstract
Survival of genotoxic stress is essential to the survival of any organism. The DNA damage checkpoint (DDC) is a regulatory system that controls the cellular response to DNA damage. Dysfunction of components of the mammalian DDC, such as ATM, hChk2, p53, and BRCAl, correlates with increased cancer risk. DDC mechanisms are conserved; in Saccharomyces cerevisiae, ATM-family kinase MEC1 is required for the DDC, as are hChk2- homolog Rad53 and BRCAl-like Rad9. The powerful genetic and biochemical techniques available in the S. cerevisiae system present it as an ideal model organism in which to study the conserved DDC mechanisms. Mec1-dependent phosphorylation of Rad53 correlates with the propagation of the DDC signal. Mecl is also required for the DDC-dependent phosphorylation of Rad9, and Rad53 interacts with phosphorylated Rad9, suggesting that Rad9 acts as an adaptor for the DDC signaling pathway. A goal of this work is to characterize physical and catalytic interactions between Mecl, Rad53, and Rad9. As reported herein, I determined the contribution of Mecl consensus phosphorylation sites within Rad9 to the functions of Rad9 in the regulation of Rad53 and the DNA damage checkpoint pathway. Further, I demonstrated the direct binding of Rad53 FHA domains to phosphorylated Rad9 peptide in vitro.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2001
- Accession Number
- ADB282776
Entities
People
- David F. Stern
- Marc F. Schwartz
Organizations
- Yale University