PMID:25138221

Citation	Yadav, T, Carrasco, B, Serrano, E and Alonso, JC (2014) Roles of Bacillus subtilis DprA and SsbA in RecA-mediated genetic recombination. J. Biol. Chem. 289:27640-52
Abstract	Bacillus subtilis competence-induced RecA, SsbA, SsbB, and DprA are required to internalize and to recombine single-stranded (ss) DNA with homologous resident duplex. RecA, in the ATP · Mg(2+)-bound form (RecA · ATP), can nucleate and form filament onto ssDNA but is inactive to catalyze DNA recombination. We report that SsbA or SsbB bound to ssDNA blocks the RecA filament formation and fails to activate recombination. DprA facilitates RecA filamentation; however, the filaments cannot engage in DNA recombination. When ssDNA was preincubated with SsbA, but not SsbB, DprA was able to activate DNA strand exchange dependent on RecA · ATP. This work demonstrates that RecA · ATP, in concert with SsbA and DprA, catalyzes DNA strand exchange, and SsbB is an accessory factor in the reaction. In contrast, RecA · dATP efficiently catalyzes strand exchange even in the absence of single-stranded binding proteins or DprA, and addition of the accessory factors marginally improved it. We proposed that the RecA-bound nucleotide (ATP and to a lesser extent dATP) might dictate the requirement for accessory factors.
Links	PubMed PMC4183802 Online version:10.1074/jbc.M114.577924
Keywords	Bacillus subtilis/enzymology; Bacillus subtilis/genetics; Bacillus subtilis/metabolism; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA, Single-Stranded/genetics; DNA, Single-Stranded/metabolism; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Membrane Proteins/genetics; Membrane Proteins/metabolism; Rec A Recombinases/genetics; Rec A Recombinases/metabolism; Recombination, Genetic
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BACIU:A0A0D5D509		GO:0009294: DNA mediated transformation	IDA: Inferred from Direct Assay:	P	The authors purified DprA, RecA, SsbA, and SsbB. They then measured the ssDNA-dependent dATP hydrolysis activity of RecA after the ssDNA was coated with nothing, DprA, SsbA and SsbB, or SsbA and SsbB followed by DprA. It was found that DprA facilitated RecA nucleation onto SSB coated ssDNA and DNA strand annealing. Experimentation with RecA in the presence of DprA also showed that DprA assisted in DNA strand exchange. In addition, it was found that DprA was important in plasmid transformation as plasmid transformation was impaired in ΔdprA ΔrecA mutant cells.	complete CACAO 11563

References

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