Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins.
The DMC1 protein, a meiosis-specific DNA recombinase, catalyzes strand exchange between homologous chromosomes. In rice, two Dmc1 genes, Dmc1A and Dmc1B, have been reported. Although the Oryza sativa DMC1A protein has been partially characterized, however the biochemical properties of the DMC1B protein have not been defined. In the present study, we expressed the Oryza sativa DMC1A and DMC1B proteins in bacteria and purified them. The purified DMC1A and DMC1B proteins formed helical filaments along single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), and promoted robust strand exchange between ssDNA and dsDNA over five thousand base pairs in the presence of RPA, as a co-factor. The DMC1A and DMC1B proteins also promoted strand exchange in the absence of RPA with long DNA substrates containing several thousand base pairs. In contrast, the human DMC1 protein strictly required RPA to promote strand exchange with these long DNA substrates. The strand-exchange activity of the Oryza sativa DMC1A protein was much higher than that of the DMC1B protein. Consistently, the DNA-binding activity of the DMC1A protein was higher than that of the DMC1B protein. These biochemical differences between the DMC1A and DMC1B proteins may provide important insight into their functional differences during meiosis in rice.
Sakane
Isao
I
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 513 Wasedatsurumaki-cho, Shinjuku-ku, Tokyo 162-0041, Japan.
Kamataki
Chiaki
C
Takizawa
Yoshimasa
Y
Nakashima
Marina
M
Toki
Seiichi
S
Ichikawa
Hiroaki
H
Ikawa
Shukuko
S
Shibata
Takehiko
T
Kurumizaka
Hitoshi
H
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D004247:DNA / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D010940:Plant Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D045522:Recombinases / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D016415:Sequence Alignment
D010940:Plant Proteins; D045522:Recombinases; D004247:DNA; D000251:Adenosine Triphosphatases