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Title: Electron microscopic single particle analysis of a tetrameric RuvA/RuvB/Holliday junction DNA complex

Abstract

During the late stage of homologous recombination in prokaryotes, RuvA binds to the Holliday junction intermediate and executes branch migration in association with RuvB. The RuvA subunits form two distinct complexes with the Holliday junction: complex I with the single RuvA tetramer on one side of the four way junction DNA, and complex II with two tetramers on both sides. To investigate the functional roles of complexes I and II, we mutated two residues of RuvA (L125D and E126K) to prevent octamer formation. An electron microscopic analysis indicated that the mutant RuvA/RuvB/Holliday junction DNA complex formed the characteristic tripartite structure, with only one RuvA tetramer bound to one side of the Holliday junction, demonstrating the unexpected stability of this complex. The novel bent images of the complex revealed an intriguing morphological similarity to the structure of SV40 large T antigen, which belongs to the same AAA+ family as RuvB.

Authors:
 [1];  [2];  [3];  [4];  [5];  [2]
  1. Nagahama Institute of Bio-Science and Technology, 1266 Tamura-cho, Nagahama, Shiga 526-0829 (Japan)
  2. Japan
  3. Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531 (Japan)
  4. Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871 (Japan)
  5. The Takara-Bio Endowed Division, Institute for Protein Research, Osaka University, 6-2-3 Furuedai, Suita, Osaka 565-0874 (Japan)
Publication Date:
OSTI Identifier:
21043571
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 365; Journal Issue: 2; Other Information: DOI: 10.1016/j.bbrc.2007.10.165; PII: S0006-291X(07)02341-8; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIGENS; COMPUTERS; DNA; ELECTRON MICROSCOPY; IMAGE PROCESSING; IMAGES; MUTANTS; PROTEINS; RECOMBINATION

Citation Formats

Mayanagi, Kouta, Takara-Bio Endowed Division, Institute for Protein Research, Osaka University, 6-2-3 Furuedai, Suita, Osaka 565-0874, BIRD, JST, Fujiwara, Yoshie, Miyata, Tomoko, Morikawa, Kosuke, and CREST, JST. Electron microscopic single particle analysis of a tetrameric RuvA/RuvB/Holliday junction DNA complex. United States: N. p., 2008. Web. doi:10.1016/j.bbrc.2007.10.165.
Mayanagi, Kouta, Takara-Bio Endowed Division, Institute for Protein Research, Osaka University, 6-2-3 Furuedai, Suita, Osaka 565-0874, BIRD, JST, Fujiwara, Yoshie, Miyata, Tomoko, Morikawa, Kosuke, & CREST, JST. Electron microscopic single particle analysis of a tetrameric RuvA/RuvB/Holliday junction DNA complex. United States. doi:10.1016/j.bbrc.2007.10.165.
Mayanagi, Kouta, Takara-Bio Endowed Division, Institute for Protein Research, Osaka University, 6-2-3 Furuedai, Suita, Osaka 565-0874, BIRD, JST, Fujiwara, Yoshie, Miyata, Tomoko, Morikawa, Kosuke, and CREST, JST. Fri . "Electron microscopic single particle analysis of a tetrameric RuvA/RuvB/Holliday junction DNA complex". United States. doi:10.1016/j.bbrc.2007.10.165.
@article{osti_21043571,
title = {Electron microscopic single particle analysis of a tetrameric RuvA/RuvB/Holliday junction DNA complex},
author = {Mayanagi, Kouta and Takara-Bio Endowed Division, Institute for Protein Research, Osaka University, 6-2-3 Furuedai, Suita, Osaka 565-0874 and BIRD, JST and Fujiwara, Yoshie and Miyata, Tomoko and Morikawa, Kosuke and CREST, JST},
abstractNote = {During the late stage of homologous recombination in prokaryotes, RuvA binds to the Holliday junction intermediate and executes branch migration in association with RuvB. The RuvA subunits form two distinct complexes with the Holliday junction: complex I with the single RuvA tetramer on one side of the four way junction DNA, and complex II with two tetramers on both sides. To investigate the functional roles of complexes I and II, we mutated two residues of RuvA (L125D and E126K) to prevent octamer formation. An electron microscopic analysis indicated that the mutant RuvA/RuvB/Holliday junction DNA complex formed the characteristic tripartite structure, with only one RuvA tetramer bound to one side of the Holliday junction, demonstrating the unexpected stability of this complex. The novel bent images of the complex revealed an intriguing morphological similarity to the structure of SV40 large T antigen, which belongs to the same AAA+ family as RuvB.},
doi = {10.1016/j.bbrc.2007.10.165},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 2,
volume = 365,
place = {United States},
year = {2008},
month = {1}
}