Structure-function analysis of the three domains of RuvB DNA motor protein.
RuvB protein forms two hexameric rings that bind to the RuvA tetramer at DNA Holliday junctions. The RuvAB complex utilizes the energy of ATP hydrolysis to promote branch migration of Holliday junctions. The crystal structure of RuvB from Thermus thermophilus (Tth) HB8 showed that each RuvB monomer has three domains (N, M, and C). This study is a structure-function analysis of the three domains of RuvB. The results show that domain N is involved in RuvA-RuvB and RuvB-RuvB subunit interactions, domains N and M are required for ATP hydrolysis and ATP binding-induced hexamer formation, and domain C plays an essential role in DNA binding. The side chain of Arg-318 is essential for DNA binding and may directly interact with DNA. The data also provide evidence that coordinated ATP-dependent interactions between domains N, M, and C play an essential role during formation of the RuvAB Holliday junction ternary complex.
Ohnishi
Takayuki
T
Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
Hishida
Takashi
T
Harada
Yoshie
Y
Iwasaki
Hiroshi
H
Shinagawa
Hideo
H
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D004260:DNA Repair; D045566:DNA, Cruciform; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000378:metabolism; D005990:Glycerol / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D013696:Temperature; D016963:Thermus thermophilus / Q000378:metabolism; D013997:Time Factors; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D045566:DNA, Cruciform; D046911:Macromolecular Substances; C061260:RuvB protein, Bacteria; D000255:Adenosine Triphosphate; D004247:DNA; D000251:Adenosine Triphosphatases; D005990:Glycerol