PMID:11927569

Citation	Bochkareva, E, Korolev, S, Lees-Miller, SP and Bochkarev, A (2002) Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA. EMBO J. 21:1855-63
Abstract	The human single-stranded DNA-binding protein, replication protein A (RPA) binds DNA in at least two different modes: initial [8-10 nucleotides (nt)] and stable ( approximately 30 nt). Switching from 8 to 30 nt mode is associated with a large conformational change. Here we report the 2.8 A structure of the RPA trimerization core comprising the C-terminal DNA-binding domain of subunit RPA70 (DBD-C), the central DNA-binding domain of subunit RPA32 (DBD-D) and the entire RPA14 subunit. All three domains are built around a central oligonucleotide/oligosaccharide binding (OB)-fold and flanked by a helix at the C-terminus. Trimerization is mediated by three C-terminal helices arranged in parallel. The OB-fold of DBD-C possesses unique structural features; embedded zinc ribbon and helix-turn-helix motifs. Using time-resolved proteolysis with trypsin, we demonstrate that the trimerization core does not contribute to the binding with substrates of 10 nt, but interacts with oligonucleotides of 24 nt. Taken together, our data indicate that switching from 8-10 to 30 nt mode is mediated by DNA binding with the trimerization core.
Links	PubMed PMC125950 Online version:10.1093/emboj/21.7.1855
Keywords	Amino Acid Sequence; Binding Sites; DNA Replication; DNA, Single-Stranded/chemistry; DNA-Binding Proteins/chemistry; Models, Molecular; Molecular Sequence Data; Protein Structure, Tertiary; Replication Protein A
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:RFA2	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P27694	F	occurs_in:(GO:0005662)	Seeded From UniProt	complete
HUMAN:RFA1	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P15927	F	occurs_in:(GO:0005662)	Seeded From UniProt	complete
HUMAN:RFA1	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P35244	F	occurs_in:(GO:0005662)	Seeded From UniProt	complete
HUMAN:RFA3	part_of	GO:0005662: DNA replication factor A complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:RFA3	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P27694	F	occurs_in:(GO:0005662)	Seeded From UniProt	complete
HUMAN:RFA1	part_of	GO:0005662: DNA replication factor A complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:RFA2	part_of	GO:0005662: DNA replication factor A complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete

References

See Help:References for how to manage references in GONUTS.