Domain analysis of an archaeal RadA protein for the strand exchange activity.
Archaeal RadA, like eukaryotic Rad51 and bacterial RecA, promotes strand exchange between DNA strands with homologous sequences in vitro and is believed to participate in the homologous recombination in cells. The amino acid sequences of the archaeal RadA proteins are more similar to the eukaryotic Rad51s rather than the bacterial RecAs, and the N-terminal region containing domain I is conserved among the RadA and Rad51 proteins but is absent from RecA. To understand the structure-function relationship of RadA, we divided the RadA protein from Pyrococcus furiosus into two parts, the N-terminal one-third (RadA-n) and the residual C-terminal two-thirds (RadA-c), the latter of which contains the central core domain (domain II) of the RecA/Rad51 family proteins. RadA-c had the DNA-dependent ATPase activity and the strand exchange activity by itself, although much weaker (10%) than that of the intact RadA. These activities of RadA-c were restored to 60% of those of RadA by addition of RadA-n, indicating that the proper active structure of RadA was reconstituted in vitro. These results suggest that the basic activities of the RecA/Rad51 family proteins for homologous recombination are derived from domain II, and the N-terminal region may help to enhance the catalytic efficiencies.
Komori
K
K
Departments of Molecular Biology, Structural Biology, and Bioinformatics, Biomolecular Engineering Research Institute, Suita, Osaka 565-0874, Japan.
Miyata
T
T
Daiyasu
H
H
Toh
H
H
Shinagawa
H
H
Ishino
a Y
aY
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D019843:Archaeal Proteins; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D020027:Pyrococcus furiosus / Q000235:genetics*; D051135:Rad51 Recombinase; D011995:Recombination, Genetic; D013329:Structure-Activity Relationship
D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; C117252:RadA protein, archaeal; D004247:DNA; D051135:Rad51 Recombinase; D000251:Adenosine Triphosphatases