TY - JOUR
T1 - Structural basis of BACH1 phosphopeptide recognition by BRCA1 tandem BRCT domains
AU - Botuyan, Maria Victoria E.
AU - Nominé, Yves
AU - Yu, Xiaochun
AU - Juranic, Nenad
AU - Macura, Slobodan
AU - Chen, Junjie
AU - Mer, Georges
N1 - Funding Information:
We gratefully acknowledge Drs. Gerard Kroon, Micah Gearhart, and Dan Garrett for help with NMR analysis software, Drs. Walter Chazin, Shibani Bhattacharya, and Jaison Jacobs for pulse programs, Drs. David Waugh and David LeMaster for providing auxotrophic bacterial strains, and Dr. Ramakrishna Vadrevu for advice on selective labeling. This work is supported by grants from the National Institutes of Health to J.C. (RO1 CA89239) and to G.M. (RO1 CA109449). J.C. is a recipient of a DOD breast cancer career development award (DAMD17-02-1-0472).
PY - 2004/7
Y1 - 2004/7
N2 - BRCT tandem domains, found in many proteins involved in DNA damage checkpoint and DNA repair pathways, were recently shown to be phosphopeptide binding motifs. Using solution nuclear magnetic resonance (NMR) spectroscopy and mutational analysis, we have characterized the interaction of BRCA1-BRCT domains with a phosphoserine-containing peptide derived from the DNA repair helicase BACH1. We show that a phenylalanine in the +3 position from the phosphoserine of BACH1 is bound to a conserved hydrophobic pocket formed between the two BRCT domains and that recognition of the phosphate group is mediated by lysine and serine side chains from the amino-terminal BRCT domain. Mutations that prevent phosphopeptide binding abolish BRCA1 function in DNA damage-induced checkpoint control. Our NMR data also reveal a dynamic interaction between BRCA1-BRCT and BACH1, where the bound phosphopeptide exists as an equilibrium of two conformations and where BRCA1-BRCT undergoes a transition to a more rigid conformation upon peptide binding.
AB - BRCT tandem domains, found in many proteins involved in DNA damage checkpoint and DNA repair pathways, were recently shown to be phosphopeptide binding motifs. Using solution nuclear magnetic resonance (NMR) spectroscopy and mutational analysis, we have characterized the interaction of BRCA1-BRCT domains with a phosphoserine-containing peptide derived from the DNA repair helicase BACH1. We show that a phenylalanine in the +3 position from the phosphoserine of BACH1 is bound to a conserved hydrophobic pocket formed between the two BRCT domains and that recognition of the phosphate group is mediated by lysine and serine side chains from the amino-terminal BRCT domain. Mutations that prevent phosphopeptide binding abolish BRCA1 function in DNA damage-induced checkpoint control. Our NMR data also reveal a dynamic interaction between BRCA1-BRCT and BACH1, where the bound phosphopeptide exists as an equilibrium of two conformations and where BRCA1-BRCT undergoes a transition to a more rigid conformation upon peptide binding.
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U2 - 10.1016/j.str.2004.06.002
DO - 10.1016/j.str.2004.06.002
M3 - Article
C2 - 15242590
AN - SCOPUS:3142608985
SN - 0969-2126
VL - 12
SP - 1137
EP - 1146
JO - Structure
JF - Structure
IS - 7
ER -