Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro.
The human testis Rad51 protein, a structural homolog of E. coli RecA, binds single- and double-stranded DNA and exhibits DNA-dependent ATPase activity. Using circular ssDNA and linear dsDNA (3.0 kb in length), we demonstrate that hRad51 promotes homologous pairing and strand exchange reactions in vitro. Joint molecule formation was dependent upon ATP hydrolysis and DNA homology and was stimulated by the single-strand DNA-binding protein RP-A. In these reactions, the 5' terminus of the complementary strand of the linear duplex was efficiently transferred to the ssDNA. However, under standard conditions, extensive strand exchange was not observed. These results establish hRad51 as a functional homolog of RecA, but indicate that the human protein and its bacterial counterpart differ in their ability to promote extensive strand transfer. It is proposed that hRad51 mediates homology recognition and initiates strand exchange, but that extensive heteroduplex formation in higher organisms requires the actions of additional proteins.
Baumann
P
P
Imperial Cancer Research Fund, South Mimms, Herts, United Kingdom.
Benson
F E
FE
West
S C
SC
D000255:Adenosine Triphosphate / Q000378:metabolism*; D011232:Chemical Precipitation; D004247:DNA / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism; D004270:DNA, Circular; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D008297:Male; D009690:Nucleic Acid Conformation; D009698:Nucleoproteins / Q000378:metabolism; D011485:Protein Binding; D051135:Rad51 Recombinase; D011938:Rec A Recombinases / Q000235:genetics; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013095:Spermidine; D013737:Testis / Q000737:chemistry
D004270:DNA, Circular; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009698:Nucleoproteins; D000255:Adenosine Triphosphate; D004247:DNA; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D011938:Rec A Recombinases; D011088:DNA Ligases; D013095:Spermidine