10779/crick.12652421.v1
Amy L Cherry
Amy L
Cherry
Timothy J Nott
Timothy J
Nott
Geoffrey Kelly
Geoffrey
Kelly
Stuart L Rulten
Stuart L
Rulten
Keith W Caldecott
Keith W
Caldecott
Stephen J Smerdon
Stephen J
Smerdon
Versatility in phospho-dependent molecular recognition of the XRCC1 and XRCC4 DNA-damage scaffolds by aprataxin-family FHA domains
The Francis Crick Institute
2020
DNA break repair
DNA-damage signalling
FHA domain
Amino Acid Sequence
Binding Sites
Casein Kinase II
Crystallography, X-Ray
DNA Damage
DNA Repair
DNA Repair Enzymes
DNA-(Apurinic or Apyrimidinic Site) Lyase
DNA-Binding Proteins
Humans
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
Nuclear Proteins
Phosphorylation
Phosphotransferases (Alcohol Group Acceptor)
Poly-ADP-Ribose Binding Proteins
Protein Structure, Tertiary
X-ray Repair Cross Complementing Protein 1
Smerdon U117584228
Developmental Biology
0601 Biochemistry and Cell Biology
2020-07-14 13:25:00
Journal contribution
https://crick.figshare.com/articles/journal_contribution/Versatility_in_phospho-dependent_molecular_recognition_of_the_XRCC1_and_XRCC4_DNA-damage_scaffolds_by_aprataxin-family_FHA_domains/12652421
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.