RecX protein abrogates ATP hydrolysis and strand exchange promoted by RecA: insights into negative regulation of homologous recombination.
In many eubacteria, coexpression of recX with recA is essential for attenuation of the deleterious effects of recA overexpression; however, the molecular mechanism has remained enigmatic. Here, we show that Mycobacterium tuberculosis RecX binds directly to M. tuberculosis RecA as well as M. smegmatis and E. coli RecA proteins in vivo and in vitro, but not single-stranded DNA binding protein. The direct association of RecX with RecA failed to regulate the specificity or extent of binding of RecA either to DNA or ATP, ligands that are central to activation of its functions. Significantly, RecX severely impeded ATP hydrolysis and the generation of heteroduplex DNA promoted by homologous, as well as heterologous, RecA proteins. These findings reveal a mode of negative regulation of RecA, and imply that RecX might act as an anti-recombinase to quell inappropriate recombinational repair during normal DNA metabolism.
Venkatesh
R
R
Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
Ganesh
N
N
Guhan
N
N
Reddy
M Sreedhar
MS
Chandrasekhar
T
T
Muniyappa
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D004269:DNA, Bacterial / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D006868:Hydrolysis; D008954:Models, Biological; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D029968:Escherichia coli Proteins; C430523:RecX protein, Xanthomonas campestris; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases