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Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158
DNA replication initiation is a vital and tightly regulated step in all replicons and requires an initiator factor that specifically recognizes the DNA replication origin and starts replication. RepB from the promiscuous streptococcal plasmid pMV158 is a hexameric ring protein evolutionary related t...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4753449/ https://www.ncbi.nlm.nih.gov/pubmed/26875695 http://dx.doi.org/10.1038/srep20915 |
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author | Boer, D. Roeland Ruiz-Masó, José Angel Rueda, Manuel Petoukhov, Maxim V. Machón, Cristina Svergun, Dmitri I. Orozco, Modesto del Solar, Gloria Coll, Miquel |
author_facet | Boer, D. Roeland Ruiz-Masó, José Angel Rueda, Manuel Petoukhov, Maxim V. Machón, Cristina Svergun, Dmitri I. Orozco, Modesto del Solar, Gloria Coll, Miquel |
author_sort | Boer, D. Roeland |
collection | PubMed |
description | DNA replication initiation is a vital and tightly regulated step in all replicons and requires an initiator factor that specifically recognizes the DNA replication origin and starts replication. RepB from the promiscuous streptococcal plasmid pMV158 is a hexameric ring protein evolutionary related to viral initiators. Here we explore the conformational plasticity of the RepB hexamer by i) SAXS, ii) sedimentation experiments, iii) molecular simulations and iv) X-ray crystallography. Combining these techniques, we derive an estimate of the conformational ensemble in solution showing that the C-terminal oligomerisation domains of the protein form a rigid cylindrical scaffold to which the N-terminal DNA-binding/catalytic domains are attached as highly flexible appendages, featuring multiple orientations. In addition, we show that the hinge region connecting both domains plays a pivotal role in the observed plasticity. Sequence comparisons and a literature survey show that this hinge region could exists in other initiators, suggesting that it is a common, crucial structural element for DNA binding and manipulation. |
format | Online Article Text |
id | pubmed-4753449 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-47534492016-02-23 Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 Boer, D. Roeland Ruiz-Masó, José Angel Rueda, Manuel Petoukhov, Maxim V. Machón, Cristina Svergun, Dmitri I. Orozco, Modesto del Solar, Gloria Coll, Miquel Sci Rep Article DNA replication initiation is a vital and tightly regulated step in all replicons and requires an initiator factor that specifically recognizes the DNA replication origin and starts replication. RepB from the promiscuous streptococcal plasmid pMV158 is a hexameric ring protein evolutionary related to viral initiators. Here we explore the conformational plasticity of the RepB hexamer by i) SAXS, ii) sedimentation experiments, iii) molecular simulations and iv) X-ray crystallography. Combining these techniques, we derive an estimate of the conformational ensemble in solution showing that the C-terminal oligomerisation domains of the protein form a rigid cylindrical scaffold to which the N-terminal DNA-binding/catalytic domains are attached as highly flexible appendages, featuring multiple orientations. In addition, we show that the hinge region connecting both domains plays a pivotal role in the observed plasticity. Sequence comparisons and a literature survey show that this hinge region could exists in other initiators, suggesting that it is a common, crucial structural element for DNA binding and manipulation. Nature Publishing Group 2016-02-15 /pmc/articles/PMC4753449/ /pubmed/26875695 http://dx.doi.org/10.1038/srep20915 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Boer, D. Roeland Ruiz-Masó, José Angel Rueda, Manuel Petoukhov, Maxim V. Machón, Cristina Svergun, Dmitri I. Orozco, Modesto del Solar, Gloria Coll, Miquel Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title | Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title_full | Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title_fullStr | Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title_full_unstemmed | Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title_short | Conformational plasticity of RepB, the replication initiator protein of promiscuous streptococcal plasmid pMV158 |
title_sort | conformational plasticity of repb, the replication initiator protein of promiscuous streptococcal plasmid pmv158 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4753449/ https://www.ncbi.nlm.nih.gov/pubmed/26875695 http://dx.doi.org/10.1038/srep20915 |
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