Activities of human recombination protein Rad51.
Homologous pairing and strand exchange, which are catalyzed by Escherichia coli RecA protein, are central to homologous recombination. Homologs of this protein are found in eukaryotes; however, little has been reported on the recombinase activities of the mammalian homologs, including the human protein, denoted HsRad51. For the studies described here, we purified HsRad51 form E. coli. Although the activities of HsRad51 and RecA were qualitatively similar in the presence of ATP, there were also striking differences. The stoichiometry of binding to DNA and the rate of renaturation of complementary strands were similar for the two proteins, but rates of ATP hydrolysis, homologous pairing, and subsequent strand exchange promoted by HsRad51 were less than 1/10 those of RecA. In addition, HsRad51 bound gamma-thio-ATP and formed stable presynaptic complexes that promoted renaturation as rapidly as RecA, but the recombinant human protein catalyzed neither strand exchange nor homologous pairing of a single strand with duplex DNA in the presence of the ATP analog. By contrast, RecA promoted both of the latter reactions in control experiments. These observations suggest that among RecA-like proteins, HsRad51 may be a variant in which homologous pairing and strand exchange are more closely linked to the hydrolysis of ATP.
Gupta
R C
RC
Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA.
Bazemore
L R
LR
Golub
E I
EI
Radding
C M
CM
D000255:Adenosine Triphosphate / Q000378:metabolism; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004735:Energy Transfer; D006801:Humans; D008969:Molecular Sequence Data; D009695:Nucleic Acid Renaturation; D009841:Oligonucleotides / Q000737:chemistry; D011485:Protein Binding; D051135:Rad51 Recombinase; D011995:Recombination, Genetic; D013050:Spectrometry, Fluorescence
D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D000255:Adenosine Triphosphate; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D003850:Deoxyribonuclease I