PMID:21359151

Citation	Stohl, EA, Gruenig, MC, Cox, MM and Seifert, HS (2011) Purification and characterization of the RecA protein from Neisseria gonorrhoeae. PLoS ONE 6:e17101
Abstract	The strict human pathogen Neisseria gonorrhoeae is the only causative agent of the sexually transmitted infection gonorrhea. The recA gene from N. gonorrhoeae is essential for DNA repair, natural DNA transformation, and pilin antigenic variation, all processes that are important for the pathogenesis and persistence of N. gonorrhoeae in the human population. To understand the biochemical features of N. gonorrhoeae RecA (RecA(Ng)), we overexpressed and purified the RecA(Ng) and SSB(Ng) proteins and compared their activities to those of the well-characterized E. coli RecA and SSB proteins in vitro. We observed that RecA(Ng) promoted more strand exchange at early time points than RecA(Ec) through DNA homologous substrates, and exhibited the highest ATPase activity of any RecA protein characterized to date. Further analysis of this robust ATPase activity revealed that RecA(Ng) is more efficient at displacing SSB from ssDNA and that RecA(Ng) shows higher ATPase activity during strand exchange than RecA(Ec). Using substrates created to mimic the cellular processes of DNA transformation and pilin antigenic variation we observed that RecA(Ec) catalyzed more strand exchange through a 100 bp heterologous insert, but that RecA(Ng) catalyzed more strand exchange through regions of microheterology. Together, these data suggest that the processes of ATP hydrolysis and DNA strand exchange may be coupled differently in RecA(Ng) than in RecA(Ec). This difference may explain the unusually high ATPase activity observed for RecA(Ng) with the strand exchange activity between RecA(Ng) and RecA(Ec) being more similar.
Links	PubMed PMC3040777 Online version:10.1371/journal.pone.0017101
Keywords	Amino Acid Sequence; Bacterial Proteins/metabolism; Base Pairing/genetics; Base Sequence; Cloning, Molecular; DNA, Single-Stranded/metabolism; Humans; Magnesium/pharmacology; Molecular Sequence Data; Neisseria gonorrhoeae/chemistry; Neisseria gonorrhoeae/genetics; Neisseria gonorrhoeae/metabolism; Rec A Recombinases/genetics; Rec A Recombinases/isolation & purification; Rec A Recombinases/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Serine Endopeptidases/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
NEIGO:RECA	enables	GO:0016887: ATPase activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
NEIGO:RECA		GO:0006310: DNA recombination	ECO:0000247:	UniProtKB:P0A7G6	P		Figure 1 shows an amino acid sequence alignment of recA from N. gonorrhoeae aligned to E.coli	complete CACAO 8723
NEIGO:RECA		GO:0016887: ATPase activity	ECO:0000314:		F		Figure 5 measures the amount of ATP hydrolyzed by RecA over time during strand exchange	complete CACAO 8724
NEIG2:B4RKU5		GO:0006260: DNA replication	ECO:0000247:	UniProtKB:P0AGE0	P		Figure 1 shows the amino acid sequence alignment between SSB of Neisseria gonorrhoeae and SSB in E.coli	complete CACAO 8725

References

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