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The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA
The MgaSpn transcriptional regulator contributes to the virulence of Streptococcus pneumoniae. It is thought to be a member of the Mga/AtxA family of global regulators. MgaSpn was shown to activate in vivo the P1623B promoter, which is divergent from the promoter (Pmga) of its own gene. This activat...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737547/ https://www.ncbi.nlm.nih.gov/pubmed/23723245 http://dx.doi.org/10.1093/nar/gkt445 |
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author | Solano-Collado, Virtu Lurz, Rudi Espinosa, Manuel Bravo, Alicia |
author_facet | Solano-Collado, Virtu Lurz, Rudi Espinosa, Manuel Bravo, Alicia |
author_sort | Solano-Collado, Virtu |
collection | PubMed |
description | The MgaSpn transcriptional regulator contributes to the virulence of Streptococcus pneumoniae. It is thought to be a member of the Mga/AtxA family of global regulators. MgaSpn was shown to activate in vivo the P1623B promoter, which is divergent from the promoter (Pmga) of its own gene. This activation required a 70-bp region (PB activation region) located between both promoters. In this work, we purified an untagged form of the MgaSpn protein, which formed dimers in solution. By gel retardation and footprinting assays, we analysed the binding of MgaSpn to linear double-stranded DNAs. MgaSpn interacted with the PB activation region when it was placed at internal position on the DNA. However, when it was positioned at one DNA end, MgaSpn recognized preferentially the Pmga promoter placed at internal position. In both cases, and on binding to the primary site, MgaSpn spread along the adjacent DNA regions generating multimeric protein–DNA complexes. When both MgaSpn-binding sites were located at internal positions on longer DNAs, electron microscopy experiments demonstrated that the PB activation region was the preferred target. DNA molecules totally or partially covered by MgaSpn were also visualized. Our results suggest that MgaSpn might recognize particular DNA conformations to achieve DNA-binding specificity. |
format | Online Article Text |
id | pubmed-3737547 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-37375472013-08-08 The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA Solano-Collado, Virtu Lurz, Rudi Espinosa, Manuel Bravo, Alicia Nucleic Acids Res Molecular Biology The MgaSpn transcriptional regulator contributes to the virulence of Streptococcus pneumoniae. It is thought to be a member of the Mga/AtxA family of global regulators. MgaSpn was shown to activate in vivo the P1623B promoter, which is divergent from the promoter (Pmga) of its own gene. This activation required a 70-bp region (PB activation region) located between both promoters. In this work, we purified an untagged form of the MgaSpn protein, which formed dimers in solution. By gel retardation and footprinting assays, we analysed the binding of MgaSpn to linear double-stranded DNAs. MgaSpn interacted with the PB activation region when it was placed at internal position on the DNA. However, when it was positioned at one DNA end, MgaSpn recognized preferentially the Pmga promoter placed at internal position. In both cases, and on binding to the primary site, MgaSpn spread along the adjacent DNA regions generating multimeric protein–DNA complexes. When both MgaSpn-binding sites were located at internal positions on longer DNAs, electron microscopy experiments demonstrated that the PB activation region was the preferred target. DNA molecules totally or partially covered by MgaSpn were also visualized. Our results suggest that MgaSpn might recognize particular DNA conformations to achieve DNA-binding specificity. Oxford University Press 2013-08 2013-05-30 /pmc/articles/PMC3737547/ /pubmed/23723245 http://dx.doi.org/10.1093/nar/gkt445 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Molecular Biology Solano-Collado, Virtu Lurz, Rudi Espinosa, Manuel Bravo, Alicia The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title | The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title_full | The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title_fullStr | The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title_full_unstemmed | The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title_short | The pneumococcal MgaSpn virulence transcriptional regulator generates multimeric complexes on linear double-stranded DNA |
title_sort | pneumococcal mgaspn virulence transcriptional regulator generates multimeric complexes on linear double-stranded dna |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737547/ https://www.ncbi.nlm.nih.gov/pubmed/23723245 http://dx.doi.org/10.1093/nar/gkt445 |
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