Structure of the DNA-bound BRCA1 C-terminal region from human replication factor C p140 and model of the protein-DNA complex.
BRCA1 C-terminal domain (BRCT)-containing proteins are found widely throughout the animal and bacteria kingdoms where they are exclusively involved in cell cycle regulation and DNA metabolism. Whereas most BRCT domains are involved in protein-protein interactions, a small subset has bona fide DNA binding activity. Here, we present the solution structure of the BRCT region of the large subunit of replication factor C bound to DNA and a model of the structure-specific complex with 5'-phosphorylated double-stranded DNA. The replication factor C BRCT domain possesses a large basic patch on one face, which includes residues that are structurally conserved and ligate the phosphate in phosphopeptide binding BRCT domains. An extra alpha-helix at the N terminus, which is required for DNA binding, inserts into the major groove and makes extensive contacts to the DNA backbone. The model of the protein-DNA complex suggests 5'-phosphate recognition by the BRCT domains of bacterial NAD(+)-dependent ligases and a nonclamp loading role for the replication factor C complex in DNA transactions.
Kobayashi
Masakazu
M
Leiden Institute of Chemistry, Leiden University, Leiden 2300RA.
Ab
Eiso
E
Department of NMR Spectroscopy, Bijvoet Center for Biomolecular Research, University of Utrecht, Utrecht 3584, Netherlands.
Bonvin
Alexander M J J
AMJJ
Department of NMR Spectroscopy, Bijvoet Center for Biomolecular Research, University of Utrecht, Utrecht 3584, Netherlands.
Siegal
Gregg
G
Leiden Institute of Chemistry, Leiden University, Leiden 2300RA. Electronic address: g.siegal@chem.leidenuniv.nl.
D000465:Algorithms; D000595:Amino Acid Sequence; D019313:BRCA1 Protein / Q000737:chemistry*; D019295:Computational Biology / Q000379:methods; D004247:DNA / Q000737:chemistry*; D004261:DNA Replication; D006801:Humans; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D051818:Replication Protein C / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid
D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D004247:DNA; D051818:Replication Protein C