Structural details of the binding of RPA and ATRIP from NMR spectroscopy and cell biology

BIOL 158

Mark R. Ehrhardt, mark.ehrhardt@vanderbilt.edu1, Heather L. Ball2, Daniel A. Mordes2, David Cortez2, and Walter J. Chazin1. (1) Center for Structural Biology, Vanderbilt University, 5140 MRB III, 465 21st Avenue South, Nashville, TN 37232, (2) Department of Biochemistry, Vanderbilt University
Phosphoinositide 3-kinase related protein kinases (PIKK) including ataxia-telangiectasia mutated (ATM), DNA dependent protein kinase (DNA-PK), and ATM and Rad3-related (ATR) coordinate cellular responses to DNA damage. DNA-PK and ATM are primarily activated by double-strand breaks. The ATR kinase, in contrast, responds to numerous forms of genotoxic stress including intrastrand cross-links, oxidative damage, and polymerase toxins. Recent data indicate that single-stranded DNA (ssDNA) coated with the single-stranded DNA-binding protein Replication Protein A (RPA) is a common intermediate responsible for activating ATR signaling in response to all of these genotoxic lesions. This RPA-ssDNA intermediate is produced via the decoupling of helicase and polymerase activities at a replication fork. It has been demonstrated that a functional domain at the N terminus of ATRIP is necessary and sufficient for interaction with RPA-ssDNA. We present here a detailed structural analysis of the primary binding interaction between RPA and ATRIP utilizing a range of biophysical techniques including NMR spectroscopy.
 

Chemistry and Metabolism
4:30 PM-6:30 PM, Tuesday, 12 September 2006 Moscone Center -- Hall D, Poster

Division of Biological Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006