Mechanisms and Components of the DNA Damage Checkpoint
Abstract
Survival of genotoxic shock is essential to the survival of any organism. The DNA damage checkpoint (DDC) controls responses to DNA damage. Dysfunction of components of the mammalian DDC pathways, such as ATM, hChk2, p53, and BRCAl, correlate with increased cancer risk. DDC mechanisms are conserved; in Saccharomyces cerevisiae, A TM-family kinase MEC1 is required for the DDC, as are hChk2-homolog Rad53 and BRCAl-like Rad9. The powerful genetic and biochemical techniques in the S. cerevisiae model present it as an ideal system in which to study conserved DDC mechanisms. DNA damage induces the MEC1-dependent phosphorylation of Rad53. MEC1 is also required for the damage-dependent phosphorylation of Rad9, and Rad53 interacts with phosphorylated Rad9, suggesting that Rad9 is an adaptor for the DDC. The goal of this work is to characterize physical and catalytic interactions between Meci, RadS3, and Rad9. As summarized herein, I identified multiple Meci phosphorylation sites within Rad9 that are induced in response to DNA damage. I determined the contribution of these phosphorylation sites to Rad9 function, including the interaction of Rad9 with Rad53. Finally, I reconstructed the interaction of phosphorylated Rad9 with in vitro, demonstrating that the Rad53 FHA domains specifically bind the phosphorylated form of Rad9 peptides.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2002
- Accession Number
- ADB286264
Entities
People
- David F. Stern
- Marc F. Schwartz
Organizations
- Yale University