Structural insights into recognition of MDC1 by TopBP1 in DNA replication checkpoint control

Charles Chung Yun Leung, Luxin Sun, Zihua Gong, Michael Burkat, Ross Edwards, Mark Assmus, Junjie Chen, J. N.Mark Glover*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Activation of the DNA replication checkpoint by the ATR kinase requires protein interactions mediated by the ATR-activating protein, TopBP1. Accumulation of TopBP1 at stalled replication forks requires the interaction of TopBP1 BRCT5 with the phosphorylated SDT repeats of the adaptor protein MDC1. Here, we present the X-ray crystal structures of the tandem BRCT4/5 domains of TopBP1 free and in complex with a MDC1 consensus pSDpT phosphopeptide. TopBP1 BRCT4/5 adopts a variant BRCT-BRCT packing interface and recognizes its target peptide in a manner distinct from that observed in previous tandem BRCT- peptide structures. The phosphate-binding pocket and positively charged residues in a variant loop in BRCT5 present an extended binding surface for the negatively charged MDC1 phosphopeptide. Mutations in this surface reduce binding affinity and recruitment of TopBP1 to γH2AX foci in cells. These studies reveal a different mode of phosphopeptide binding by BRCT domains in the DNA damage response.

Original languageEnglish (US)
Pages (from-to)1450-1459
Number of pages10
JournalStructure
Volume21
Issue number8
DOIs
StatePublished - Aug 6 2013
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Structural insights into recognition of MDC1 by TopBP1 in DNA replication checkpoint control'. Together they form a unique fingerprint.

Cite this