Versatility in phospho-dependent molecular recognition of the XRCC1 and XRCC4 DNA-damage scaffolds by aprataxin-family FHA domains

Research output: Contribution to journalArticle

Authors

  • Amy L Cherry
  • Timothy J Nott
  • Geoffrey Kelly
  • Stuart L Rulten
  • Keith W Caldecott

Colleges, School and Institutes

External organisations

  • Francis Crick Institute, Mill Hill Laboratory, The Ridgeway, Mill Hill, London NW7 1AA, UK.
  • MRC Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, UK.

Abstract

Aprataxin, aprataxin and PNKP-like factor (APLF) and polynucleotide kinase phosphatase (PNKP) are key DNA-repair proteins with diverse functions but which all contain a homologous forkhead-associated (FHA) domain. Their primary binding targets are casein kinase 2-phosphorylated forms of the XRCC1 and XRCC4 scaffold molecules which respectively coordinate single-stranded and double-stranded DNA break repair pathways. Here, we present the high-resolution X-ray structure of a complex of phosphorylated XRCC4 with APLF, the most divergent of the three FHA domain family members. This, combined with NMR and biochemical analysis of aprataxin and APLF binding to singly and multiply-phosphorylated forms of XRCC1 and XRCC4, and comparison with PNKP reveals a pattern of distinct but overlapping binding specificities that are differentially modulated by multi-site phosphorylation. Together, our data illuminate important differences between activities of the three phospho-binding domains, in spite of a close evolutionary relationship between them.

Bibliographic note

Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Details

Original languageEnglish
Pages (from-to)116-25
Number of pages10
JournalDNA Repair
Volume35
Publication statusPublished - Nov 2015

Keywords

  • Amino Acid Sequence, Binding Sites, Casein Kinase II/metabolism, Crystallography, X-Ray, DNA Damage, DNA Repair, DNA Repair Enzymes/ultrastructure, DNA-(Apurinic or Apyrimidinic Site) Lyase/chemistry, DNA-Binding Proteins/chemistry, Humans, Molecular Sequence Data, Nuclear Magnetic Resonance, Biomolecular, Nuclear Proteins/chemistry, Phosphorylation, Phosphotransferases (Alcohol Group Acceptor)/ultrastructure, Poly-ADP-Ribose Binding Proteins, Protein Structure, Tertiary, X-ray Repair Cross Complementing Protein 1