A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.

Small RNAs (sRNAs) are becoming increasingly recognized as important regulators in bacteria. To investigate the contribution of sRNA mediated regulation to virulence in Vibrio cholerae, we performed high throughput sequencing of cDNA generated from sRNA transcripts isolated from a strain ectopically...

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Autores principales:	Evan S Bradley, Kip Bodi, Ayman M Ismail, Andrew Camilli
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/3af6f42ba13c499697de0ce6189749d4
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spelling	oai:doaj.org-article:3af6f42ba13c499697de0ce6189749d42021-11-18T06:03:13ZA genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.1553-73661553-737410.1371/journal.ppat.1002126https://doaj.org/article/3af6f42ba13c499697de0ce6189749d42011-07-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21779167/pdf/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Small RNAs (sRNAs) are becoming increasingly recognized as important regulators in bacteria. To investigate the contribution of sRNA mediated regulation to virulence in Vibrio cholerae, we performed high throughput sequencing of cDNA generated from sRNA transcripts isolated from a strain ectopically expressing ToxT, the major transcriptional regulator within the virulence gene regulon. We compared this data set with ToxT binding sites determined by pulldown and deep sequencing to identify sRNA promoters directly controlled by ToxT. Analysis of the resulting transcripts with ToxT binding sites in cis revealed two sRNAs within the Vibrio Pathogenicity Island. When deletions of these sRNAs were made and the resulting strains were competed against the parental strain in the infant mouse model of V. cholerae colonization, one, TarB, displayed a variable colonization phenotype dependent on its physiological state at the time of inoculation. We identified a target of TarB as the mRNA for the secreted colonization factor, TcpF. We verified negative regulation of TcpF expression by TarB and, using point mutations that disrupted interaction between TarB and tpcF mRNA, showed that loss of this negative regulation was primarily responsible for the colonization phenotype observed in the TarB deletion mutant.Evan S BradleyKip BodiAyman M IsmailAndrew CamilliPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 7, Iss 7, p e1002126 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Evan S Bradley Kip Bodi Ayman M Ismail Andrew Camilli A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
description	Small RNAs (sRNAs) are becoming increasingly recognized as important regulators in bacteria. To investigate the contribution of sRNA mediated regulation to virulence in Vibrio cholerae, we performed high throughput sequencing of cDNA generated from sRNA transcripts isolated from a strain ectopically expressing ToxT, the major transcriptional regulator within the virulence gene regulon. We compared this data set with ToxT binding sites determined by pulldown and deep sequencing to identify sRNA promoters directly controlled by ToxT. Analysis of the resulting transcripts with ToxT binding sites in cis revealed two sRNAs within the Vibrio Pathogenicity Island. When deletions of these sRNAs were made and the resulting strains were competed against the parental strain in the infant mouse model of V. cholerae colonization, one, TarB, displayed a variable colonization phenotype dependent on its physiological state at the time of inoculation. We identified a target of TarB as the mRNA for the secreted colonization factor, TcpF. We verified negative regulation of TcpF expression by TarB and, using point mutations that disrupted interaction between TarB and tpcF mRNA, showed that loss of this negative regulation was primarily responsible for the colonization phenotype observed in the TarB deletion mutant.
format	article
author	Evan S Bradley Kip Bodi Ayman M Ismail Andrew Camilli
author_facet	Evan S Bradley Kip Bodi Ayman M Ismail Andrew Camilli
author_sort	Evan S Bradley
title	A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
title_short	A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
title_full	A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
title_fullStr	A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
title_full_unstemmed	A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.
title_sort	genome-wide approach to discovery of small rnas involved in regulation of virulence in vibrio cholerae.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/3af6f42ba13c499697de0ce6189749d4
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A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae.

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