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Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners
YabA negatively regulates initiation of DNA replication in low-GC Gram-positive bacteria. The protein exerts its control through interactions with the initiator protein DnaA and the sliding clamp DnaN. Here, we combined X-ray crystallography, X-ray scattering (SAXS), modeling and biophysical approac...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705661/ https://www.ncbi.nlm.nih.gov/pubmed/26615189 http://dx.doi.org/10.1093/nar/gkv1318 |
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author | Felicori, Liza Jameson, Katie H. Roblin, Pierre Fogg, Mark J. Garcia-Garcia, Transito Ventroux, Magali Cherrier, Mickaël V. Bazin, Alexandre Noirot, Philippe Wilkinson, Anthony J. Molina, Franck Terradot, Laurent Noirot-Gros, Marie-Françoise |
author_facet | Felicori, Liza Jameson, Katie H. Roblin, Pierre Fogg, Mark J. Garcia-Garcia, Transito Ventroux, Magali Cherrier, Mickaël V. Bazin, Alexandre Noirot, Philippe Wilkinson, Anthony J. Molina, Franck Terradot, Laurent Noirot-Gros, Marie-Françoise |
author_sort | Felicori, Liza |
collection | PubMed |
description | YabA negatively regulates initiation of DNA replication in low-GC Gram-positive bacteria. The protein exerts its control through interactions with the initiator protein DnaA and the sliding clamp DnaN. Here, we combined X-ray crystallography, X-ray scattering (SAXS), modeling and biophysical approaches, with in vivo experimental data to gain insight into YabA function. The crystal structure of the N-terminal domain (NTD) of YabA solved at 2.7 Å resolution reveals an extended α-helix that contributes to an intermolecular four-helix bundle. Homology modeling and biochemical analysis indicates that the C-terminal domain (CTD) of YabA is a small Zn-binding domain. Multi-angle light scattering and SAXS demonstrate that YabA is a tetramer in which the CTDs are independent and connected to the N-terminal four-helix bundle via flexible linkers. While YabA can simultaneously interact with both DnaA and DnaN, we found that an isolated CTD can bind to either DnaA or DnaN, individually. Site-directed mutagenesis and yeast-two hybrid assays identified DnaA and DnaN binding sites on the YabA CTD that partially overlap and point to a mutually exclusive mode of interaction. Our study defines YabA as a novel structural hub and explains how the protein tetramer uses independent CTDs to bind multiple partners to orchestrate replication initiation in the bacterial cell. |
format | Online Article Text |
id | pubmed-4705661 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-47056612016-01-11 Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners Felicori, Liza Jameson, Katie H. Roblin, Pierre Fogg, Mark J. Garcia-Garcia, Transito Ventroux, Magali Cherrier, Mickaël V. Bazin, Alexandre Noirot, Philippe Wilkinson, Anthony J. Molina, Franck Terradot, Laurent Noirot-Gros, Marie-Françoise Nucleic Acids Res Structural Biology YabA negatively regulates initiation of DNA replication in low-GC Gram-positive bacteria. The protein exerts its control through interactions with the initiator protein DnaA and the sliding clamp DnaN. Here, we combined X-ray crystallography, X-ray scattering (SAXS), modeling and biophysical approaches, with in vivo experimental data to gain insight into YabA function. The crystal structure of the N-terminal domain (NTD) of YabA solved at 2.7 Å resolution reveals an extended α-helix that contributes to an intermolecular four-helix bundle. Homology modeling and biochemical analysis indicates that the C-terminal domain (CTD) of YabA is a small Zn-binding domain. Multi-angle light scattering and SAXS demonstrate that YabA is a tetramer in which the CTDs are independent and connected to the N-terminal four-helix bundle via flexible linkers. While YabA can simultaneously interact with both DnaA and DnaN, we found that an isolated CTD can bind to either DnaA or DnaN, individually. Site-directed mutagenesis and yeast-two hybrid assays identified DnaA and DnaN binding sites on the YabA CTD that partially overlap and point to a mutually exclusive mode of interaction. Our study defines YabA as a novel structural hub and explains how the protein tetramer uses independent CTDs to bind multiple partners to orchestrate replication initiation in the bacterial cell. Oxford University Press 2016-01-08 2015-11-28 /pmc/articles/PMC4705661/ /pubmed/26615189 http://dx.doi.org/10.1093/nar/gkv1318 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Structural Biology Felicori, Liza Jameson, Katie H. Roblin, Pierre Fogg, Mark J. Garcia-Garcia, Transito Ventroux, Magali Cherrier, Mickaël V. Bazin, Alexandre Noirot, Philippe Wilkinson, Anthony J. Molina, Franck Terradot, Laurent Noirot-Gros, Marie-Françoise Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title | Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title_full | Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title_fullStr | Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title_full_unstemmed | Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title_short | Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners |
title_sort | tetramerization and interdomain flexibility of the replication initiation controller yaba enables simultaneous binding to multiple partners |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705661/ https://www.ncbi.nlm.nih.gov/pubmed/26615189 http://dx.doi.org/10.1093/nar/gkv1318 |
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