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Structure paper

TitleCryo-EM structure of the RuvAB-Holliday junction intermediate complex from .
Journal, issue, pagesFront Plant Sci, Vol. 14, Page 1139106, Year 2023
Publish dateMar 21, 2023
AuthorsXu Zhang / Zixuan Zhou / Lin Dai / Yulin Chao / Zheng Liu / Mingdong Huang / Qianhui Qu / Zhonghui Lin /
PubMed AbstractHolliday junction (HJ) is a four-way structured DNA intermediate in homologous recombination. In bacteria, the HJ-specific binding protein RuvA and the motor protein RuvB together form the RuvAB ...Holliday junction (HJ) is a four-way structured DNA intermediate in homologous recombination. In bacteria, the HJ-specific binding protein RuvA and the motor protein RuvB together form the RuvAB complex to catalyze HJ branch migration. (, Pa) is a ubiquitous opportunistic bacterial pathogen that can cause serious infection in a variety of host species, including vertebrate animals, insects and plants. Here, we describe the cryo-Electron Microscopy (cryo-EM) structure of the RuvAB-HJ intermediate complex from . The structure shows that two RuvA tetramers sandwich HJ at the junction center and disrupt base pairs at the branch points of RuvB-free HJ arms. Eight RuvB subunits are recruited by the RuvA octameric core and form two open-rings to encircle two opposite HJ arms. Each RuvB subunit individually binds a RuvA domain III. The four RuvB subunits within the ring display distinct subdomain conformations, and two of them engage the central DNA duplex at both strands with their C-terminal β-hairpins. Together with the biochemical analyses, our structure implicates a potential mechanism of RuvB motor assembly onto HJ DNA.
External linksFront Plant Sci / PubMed:37025142 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.16 - 7.02 Å
Structure data

33013

7x5a

EMDB-33013, PDB-7x5a:
CryoEM structure of RuvA-Holliday junction complex
Method: EM (single particle) / Resolution: 3.01 Å

33043

7x7p

EMDB-33043, PDB-7x7p:
CryoEM structure of dsDNA-RuvB-RuvA domain3 complex
Method: EM (single particle) / Resolution: 7.02 Å

33044

7x7q

EMDB-33044, PDB-7x7q:
CryoEM structure of RuvA-RuvB-Holliday junction complex
Method: EM (single particle) / Resolution: 7.02 Å

PDB-7x5b:
Crystal structure of RuvB
Method: X-RAY DIFFRACTION / Resolution: 2.16 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-HOH:
WATER

Source
  • pseudomonas aeruginosa pao1 (bacteria)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN/DNA / RuvA / Holliday junction / Homologous recombination / DNA damage repair / DNA BINDING PROTEIN-DNA COMPLEX / MOTOR PROTEIN / RuvB / Holliday juncition / ATP hydrolysis / MOTOR PROTEIN/DNA / MOTOR PROTEIN-DNA COMPLEX / RECOMBINATION/DNA / Branch migration / RECOMBINATION-DNA COMPLEX

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