Versatility in phospho-dependent molecular recognition of the XRCC1 and XRCC4 DNA-damage scaffolds by aprataxin-family FHA domains
Research output: Contribution to journal › Article
Colleges, School and Institutes
- 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.
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.
|Number of pages||10|
|Publication status||Published - Nov 2015|
- 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