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Sci Rep. 2015 Dec 14;5:18177. doi: 10.1038/srep18177.

Synergistic effect of ATP for RuvA-RuvB-Holliday junction DNA complex formation.

Author information

1
Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.
2
Graduate School of Biostudies, Kyoto University, Sakyo, Kyoto 606-8501, Japan.
3
CREST, Japan Science and Technology Corporation (JST), Sanbancho, Chiyoda, Tokyo 102-0075, Japan.
4
Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
5
Department of Micro Engineering, Graduate School of Technology, Kyoto University, Sakyo, Kyoto 606-8501, Japan.

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.

PMID:
26658024
PMCID:
PMC4677358
DOI:
10.1038/srep18177
[Indexed for MEDLINE]
Free PMC Article

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