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The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation
Expression of type IV pili (Tfp), filamentous appendages emanating from the bacterial surface, is indispensable for efficient neisserial transformation. Tfp pass through the secretin pore consisting of the membrane protein PilQ. PilG is a polytopic membrane protein, conserved in Gram-positive and Gr...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4527729/ https://www.ncbi.nlm.nih.gov/pubmed/26248334 http://dx.doi.org/10.1371/journal.pone.0134954 |
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author | Frye, Stephan A. Lång, Emma Beyene, Getachew Tesfaye Balasingham, Seetha V. Homberset, Håvard Rowe, Alexander D. Ambur, Ole Herman Tønjum, Tone |
author_facet | Frye, Stephan A. Lång, Emma Beyene, Getachew Tesfaye Balasingham, Seetha V. Homberset, Håvard Rowe, Alexander D. Ambur, Ole Herman Tønjum, Tone |
author_sort | Frye, Stephan A. |
collection | PubMed |
description | Expression of type IV pili (Tfp), filamentous appendages emanating from the bacterial surface, is indispensable for efficient neisserial transformation. Tfp pass through the secretin pore consisting of the membrane protein PilQ. PilG is a polytopic membrane protein, conserved in Gram-positive and Gram-negative bacteria, that is required for the biogenesis of neisserial Tfp. PilG null mutants are devoid of pili and non-competent for transformation. Here, recombinant full-length, truncated and mutated variants of meningococcal PilG were overexpressed, purified and characterized. We report that meningococcal PilG directly binds DNA in vitro, detected by both an electromobility shift analysis and a solid phase overlay assay. PilG DNA binding activity was independent of the presence of the consensus DNA uptake sequence. PilG-mediated DNA binding affinity was mapped to the N-terminus and was inactivated by mutation of residues 43 to 45. Notably, reduced meningococcal transformation of DNA in vivo was observed when PilG residues 43 to 45 were substituted by alanine in situ, defining a biologically significant DNA binding domain. N-terminal PilG also interacted with the N-terminal region of PilQ, which previously was shown to bind DNA. Collectively, these data suggest that PilG and PilQ in concert bind DNA during Tfp-mediated transformation. |
format | Online Article Text |
id | pubmed-4527729 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45277292015-08-12 The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation Frye, Stephan A. Lång, Emma Beyene, Getachew Tesfaye Balasingham, Seetha V. Homberset, Håvard Rowe, Alexander D. Ambur, Ole Herman Tønjum, Tone PLoS One Research Article Expression of type IV pili (Tfp), filamentous appendages emanating from the bacterial surface, is indispensable for efficient neisserial transformation. Tfp pass through the secretin pore consisting of the membrane protein PilQ. PilG is a polytopic membrane protein, conserved in Gram-positive and Gram-negative bacteria, that is required for the biogenesis of neisserial Tfp. PilG null mutants are devoid of pili and non-competent for transformation. Here, recombinant full-length, truncated and mutated variants of meningococcal PilG were overexpressed, purified and characterized. We report that meningococcal PilG directly binds DNA in vitro, detected by both an electromobility shift analysis and a solid phase overlay assay. PilG DNA binding activity was independent of the presence of the consensus DNA uptake sequence. PilG-mediated DNA binding affinity was mapped to the N-terminus and was inactivated by mutation of residues 43 to 45. Notably, reduced meningococcal transformation of DNA in vivo was observed when PilG residues 43 to 45 were substituted by alanine in situ, defining a biologically significant DNA binding domain. N-terminal PilG also interacted with the N-terminal region of PilQ, which previously was shown to bind DNA. Collectively, these data suggest that PilG and PilQ in concert bind DNA during Tfp-mediated transformation. Public Library of Science 2015-08-06 /pmc/articles/PMC4527729/ /pubmed/26248334 http://dx.doi.org/10.1371/journal.pone.0134954 Text en © 2015 Frye et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Frye, Stephan A. Lång, Emma Beyene, Getachew Tesfaye Balasingham, Seetha V. Homberset, Håvard Rowe, Alexander D. Ambur, Ole Herman Tønjum, Tone The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title | The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title_full | The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title_fullStr | The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title_full_unstemmed | The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title_short | The Inner Membrane Protein PilG Interacts with DNA and the Secretin PilQ in Transformation |
title_sort | inner membrane protein pilg interacts with dna and the secretin pilq in transformation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4527729/ https://www.ncbi.nlm.nih.gov/pubmed/26248334 http://dx.doi.org/10.1371/journal.pone.0134954 |
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