Cargando…
Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans
Holliday junctions (HJs) are four-way DNA structures, which are an important intermediate in the process of homologous recombination. In most bacteria, HJs are cleaved by specific nucleases called RuvC resolvases at the end of homologous recombination. Deinococcus radiodurans is an extraordinary rad...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228767/ https://www.ncbi.nlm.nih.gov/pubmed/35744678 http://dx.doi.org/10.3390/microorganisms10061160 |
_version_ | 1784734561991655424 |
---|---|
author | Qin, Chen Han, Wanchun Xu, Ying Zhao, Ye Xu, Hong Tian, Bing Wang, Liangyan Hua, Yuejin |
author_facet | Qin, Chen Han, Wanchun Xu, Ying Zhao, Ye Xu, Hong Tian, Bing Wang, Liangyan Hua, Yuejin |
author_sort | Qin, Chen |
collection | PubMed |
description | Holliday junctions (HJs) are four-way DNA structures, which are an important intermediate in the process of homologous recombination. In most bacteria, HJs are cleaved by specific nucleases called RuvC resolvases at the end of homologous recombination. Deinococcus radiodurans is an extraordinary radiation-resistant bacterium and is known as an ideal model organism for elucidating DNA repair processes. Here, we described the biochemical properties and the crystal structure of RuvC from D. radiodurans (DrRuvC). DrRuvC exhibited an RNase H fold that belonged to the retroviral integrase family. Among many DNA substrates, DrRuvC specifically bound to HJ DNA and cleaved it. In particular, Mn(2+) was the preferred bivalent metal co-factor for HJ cleavage, whereas high concentrations of Mg(2+) inhibited the binding of DrRuvC to HJ. In addition, DrRuvC was crystallized and the crystals diffracted to 1.6 Å. The crystal structure of DrRuvC revealed essential amino acid sites for cleavage and binding activities, indicating that DrRuvC was a typical resolvase with a characteristic choice for metal co-factor. |
format | Online Article Text |
id | pubmed-9228767 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92287672022-06-25 Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans Qin, Chen Han, Wanchun Xu, Ying Zhao, Ye Xu, Hong Tian, Bing Wang, Liangyan Hua, Yuejin Microorganisms Article Holliday junctions (HJs) are four-way DNA structures, which are an important intermediate in the process of homologous recombination. In most bacteria, HJs are cleaved by specific nucleases called RuvC resolvases at the end of homologous recombination. Deinococcus radiodurans is an extraordinary radiation-resistant bacterium and is known as an ideal model organism for elucidating DNA repair processes. Here, we described the biochemical properties and the crystal structure of RuvC from D. radiodurans (DrRuvC). DrRuvC exhibited an RNase H fold that belonged to the retroviral integrase family. Among many DNA substrates, DrRuvC specifically bound to HJ DNA and cleaved it. In particular, Mn(2+) was the preferred bivalent metal co-factor for HJ cleavage, whereas high concentrations of Mg(2+) inhibited the binding of DrRuvC to HJ. In addition, DrRuvC was crystallized and the crystals diffracted to 1.6 Å. The crystal structure of DrRuvC revealed essential amino acid sites for cleavage and binding activities, indicating that DrRuvC was a typical resolvase with a characteristic choice for metal co-factor. MDPI 2022-06-06 /pmc/articles/PMC9228767/ /pubmed/35744678 http://dx.doi.org/10.3390/microorganisms10061160 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Qin, Chen Han, Wanchun Xu, Ying Zhao, Ye Xu, Hong Tian, Bing Wang, Liangyan Hua, Yuejin Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title | Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title_full | Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title_fullStr | Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title_full_unstemmed | Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title_short | Structural and Functional Characterization of the Holliday Junction Resolvase RuvC from Deinococcus radiodurans |
title_sort | structural and functional characterization of the holliday junction resolvase ruvc from deinococcus radiodurans |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228767/ https://www.ncbi.nlm.nih.gov/pubmed/35744678 http://dx.doi.org/10.3390/microorganisms10061160 |
work_keys_str_mv | AT qinchen structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT hanwanchun structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT xuying structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT zhaoye structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT xuhong structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT tianbing structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT wangliangyan structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans AT huayuejin structuralandfunctionalcharacterizationofthehollidayjunctionresolvaseruvcfromdeinococcusradiodurans |