Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa
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. Pseudomonas aeruginosa (P. aeruginosa, Pa) is a ubiquitous oppor...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-03-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2023.1139106/full |
| _version_ | 1827986376313798656 |
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| author | Xu Zhang Zixuan Zhou Lin Dai Yulin Chao Zheng Liu Mingdong Huang Qianhui Qu Zhonghui Lin |
| author_facet | Xu Zhang Zixuan Zhou Lin Dai Yulin Chao Zheng Liu Mingdong Huang Qianhui Qu Zhonghui Lin |
| author_sort | Xu Zhang |
| collection | DOAJ |
| description | 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. Pseudomonas aeruginosa (P. aeruginosa, 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 P. aeruginosa. 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. |
| first_indexed | 2024-04-09T23:29:19Z |
| format | Article |
| id | doaj.art-138bd9e4d6a2419999ff5662d9e715f6 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-09T23:29:19Z |
| publishDate | 2023-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-138bd9e4d6a2419999ff5662d9e715f62023-03-21T06:12:28ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2023-03-011410.3389/fpls.2023.11391061139106Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosaXu Zhang0Zixuan Zhou1Lin Dai2Yulin Chao3Zheng Liu4Mingdong Huang5Qianhui Qu6Zhonghui Lin7College of Chemistry, Fuzhou University, Fuzhou, ChinaShanghai Stomatological Hospital, Institutes of Biomedical Science, Department of Systems Biology for Medicine, Fudan University, Shanghai, ChinaCollege of Chemistry, Fuzhou University, Fuzhou, ChinaShanghai Stomatological Hospital, Institutes of Biomedical Science, Department of Systems Biology for Medicine, Fudan University, Shanghai, ChinaKobilka Institute of Innovative Drug Discovery, School of Medicine, Chinese University of Hong Kong, Shenzhen, ChinaCollege of Chemistry, Fuzhou University, Fuzhou, ChinaShanghai Stomatological Hospital, Institutes of Biomedical Science, Department of Systems Biology for Medicine, Fudan University, Shanghai, ChinaCollege of Chemistry, Fuzhou University, Fuzhou, ChinaHolliday 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. Pseudomonas aeruginosa (P. aeruginosa, 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 P. aeruginosa. 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.https://www.frontiersin.org/articles/10.3389/fpls.2023.1139106/fullhomologous recombinationDNA damage repairHolliday junctionRuvARuvBcomplex assembly |
| spellingShingle | Xu Zhang Zixuan Zhou Lin Dai Yulin Chao Zheng Liu Mingdong Huang Qianhui Qu Zhonghui Lin Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa Frontiers in Plant Science homologous recombination DNA damage repair Holliday junction RuvA RuvB complex assembly |
| title | Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa |
| title_full | Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa |
| title_fullStr | Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa |
| title_full_unstemmed | Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa |
| title_short | Cryo-EM structure of the RuvAB-Holliday junction intermediate complex from Pseudomonas aeruginosa |
| title_sort | cryo em structure of the ruvab holliday junction intermediate complex from pseudomonas aeruginosa |
| topic | homologous recombination DNA damage repair Holliday junction RuvA RuvB complex assembly |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2023.1139106/full |
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