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Identification of neisserial DNA binding components

Neisseria meningitidis, a causative agent of meningitis and septicaemia, expresses type IV pili, a feature correlating with the uptake of exogenous DNA from the environment by natural transformation. The outer membrane complex PilQ, through which pili are extruded and retracted, has previously been...

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Autores principales: Lång, Emma, Haugen, Kristine, Fleckenstein, Burkhard, Homberset, Håvard, Frye, Stephan A., Ambur, Ole Herman, Tønjum, Tone
Formato: Texto
Lenguaje:English
Publicado: Microbiology Society 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2885667/
https://www.ncbi.nlm.nih.gov/pubmed/19246756
http://dx.doi.org/10.1099/mic.0.022640-0
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author Lång, Emma
Haugen, Kristine
Fleckenstein, Burkhard
Homberset, Håvard
Frye, Stephan A.
Ambur, Ole Herman
Tønjum, Tone
author_facet Lång, Emma
Haugen, Kristine
Fleckenstein, Burkhard
Homberset, Håvard
Frye, Stephan A.
Ambur, Ole Herman
Tønjum, Tone
author_sort Lång, Emma
collection PubMed
description Neisseria meningitidis, a causative agent of meningitis and septicaemia, expresses type IV pili, a feature correlating with the uptake of exogenous DNA from the environment by natural transformation. The outer membrane complex PilQ, through which pili are extruded and retracted, has previously been shown to bind DNA in its pore region. In order to further elucidate how DNA is transported across the membranes, we searched for DNA binding proteins within the meningococcal inner membrane. Inner membrane fractions from a panel of neisserial strains were subjected to a solid-phase overlay assay with DNA substrates, and MS was subsequently employed to identify proteins that bind DNA. A number of DNA binding components were detected, including the pilus biogenesis component PilG, the competence protein ComL, and the cell division ATP-binding protein FtsE, as well as two hypothetical proteins. The DNA binding activity of these components was not dependent on the presence of the neisserial DNA uptake sequence. Null mutants, corresponding to each of the proteins identified, were constructed to assess their phenotypes. Only mutants defective in pilus biogenesis were non-competent and non-piliated. The DNA binding activity of the pilus biogenesis components PilQ and PilG and the phenotypes of their respective null mutants suggest that these proteins are directly involved as players in natural transformation, and not only indirectly, through pilus biogenesis.
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spelling pubmed-28856672010-07-06 Identification of neisserial DNA binding components Lång, Emma Haugen, Kristine Fleckenstein, Burkhard Homberset, Håvard Frye, Stephan A. Ambur, Ole Herman Tønjum, Tone Microbiology (Reading) Microbial Pathogenicity Neisseria meningitidis, a causative agent of meningitis and septicaemia, expresses type IV pili, a feature correlating with the uptake of exogenous DNA from the environment by natural transformation. The outer membrane complex PilQ, through which pili are extruded and retracted, has previously been shown to bind DNA in its pore region. In order to further elucidate how DNA is transported across the membranes, we searched for DNA binding proteins within the meningococcal inner membrane. Inner membrane fractions from a panel of neisserial strains were subjected to a solid-phase overlay assay with DNA substrates, and MS was subsequently employed to identify proteins that bind DNA. A number of DNA binding components were detected, including the pilus biogenesis component PilG, the competence protein ComL, and the cell division ATP-binding protein FtsE, as well as two hypothetical proteins. The DNA binding activity of these components was not dependent on the presence of the neisserial DNA uptake sequence. Null mutants, corresponding to each of the proteins identified, were constructed to assess their phenotypes. Only mutants defective in pilus biogenesis were non-competent and non-piliated. The DNA binding activity of the pilus biogenesis components PilQ and PilG and the phenotypes of their respective null mutants suggest that these proteins are directly involved as players in natural transformation, and not only indirectly, through pilus biogenesis. Microbiology Society 2009-03 /pmc/articles/PMC2885667/ /pubmed/19246756 http://dx.doi.org/10.1099/mic.0.022640-0 Text en Copyright © 2009, SGM http://creativecommons.org/licenses/by/2.5/ 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 work is properly cited.
spellingShingle Microbial Pathogenicity
Lång, Emma
Haugen, Kristine
Fleckenstein, Burkhard
Homberset, Håvard
Frye, Stephan A.
Ambur, Ole Herman
Tønjum, Tone
Identification of neisserial DNA binding components
title Identification of neisserial DNA binding components
title_full Identification of neisserial DNA binding components
title_fullStr Identification of neisserial DNA binding components
title_full_unstemmed Identification of neisserial DNA binding components
title_short Identification of neisserial DNA binding components
title_sort identification of neisserial dna binding components
topic Microbial Pathogenicity
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2885667/
https://www.ncbi.nlm.nih.gov/pubmed/19246756
http://dx.doi.org/10.1099/mic.0.022640-0
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