title: Synergistic effect of ATP for RuvA-RuvB-Holliday junction DNA complex formation.
abstract: The Escherichia coli RuvB hexameric ring motor proteins, together with RuvAs, promote branch migration of Holliday junction DNA. Zero mode waveguides (ZMWs) constitute of nanosized holes and enable the visualization of a single fluorescent molecule under micromolar order of the molecules, which is applicable to characterize the formation of RuvA-RuvB-Holliday junction DNA complex. In this study, we used ZMWs and counted the number of RuvBs binding to RuvA-Holliday junction DNA complex. Our data demonstrated that different nucleotide analogs increased the amount of Cy5-RuvBs binding to RuvA-Holliday junction DNA complex in the following order: no nucleotide, ADP, ATPγS, and mixture of ADP and ATPγS. These results suggest that not only ATP binding to RuvB but also ATP hydrolysis by RuvB facilitates a stable RuvA-RuvB-Holliday junction DNA complex formation.
authors.affiliation: Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.|Graduate School of Biostudies, Kyoto University, Sakyo, Kyoto 606-8501, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Iwasa
authors.affiliation: Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.|CREST, Japan Science and Technology Corporation (JST), Sanbancho, Chiyoda, Tokyo 102-0075, Japan.
authors.forename: Yong-Woon
authors.initials: YW
authors.lastname: Han
authors.affiliation: Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Hiramatsu
authors.affiliation: Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yokota
authors.affiliation: Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.
authors.forename: Kimiko
authors.initials: K
authors.lastname: Nakao
authors.affiliation: Department of Micro Engineering, Graduate School of Technology, Kyoto University, Sakyo, Kyoto 606-8501, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Yokokawa
authors.affiliation: Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Ono
authors.affiliation: Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.|Graduate School of Biostudies, Kyoto University, Sakyo, Kyoto 606-8501, Japan.|CREST, Japan Science and Technology Corporation (JST), Sanbancho, Chiyoda, Tokyo 102-0075, Japan.
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Harada
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004265:DNA Helicases / Q000737:chemistry / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D045566:DNA, Cruciform / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D011485:Protein Binding; D021381:Protein Transport
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D045566:DNA, Cruciform; D029968:Escherichia coli Proteins; C061260:RuvB protein, Bacteria; C071059:Holliday junction DNA helicase, E coli; D000251:Adenosine Triphosphatases; D004265:DNA Helicases