The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>

ABSTRACT Nearly all virulence factors in Bordetella pertussis are activated by a master two-component system, BvgAS, composed of the sensor kinase BvgS and the response regulator BvgA. When BvgS is active, BvgA is phosphorylated (BvgA~P), and virulence-activated genes (vags) are expressed [Bvg(+) mo...

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Autores principales:	Kyung Moon, Richard P. Bonocora, David D. Kim, Qing Chen, Joseph T. Wade, Scott Stibitz, Deborah M. Hinton
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	BvgAS regulon RNA polymerase RNA-seq brpL pertussis Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0ad66eba78e94091ad2010201f7418fd
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spelling	oai:doaj.org-article:0ad66eba78e94091ad2010201f7418fd2021-11-15T15:51:51ZThe BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>10.1128/mBio.01526-172150-7511https://doaj.org/article/0ad66eba78e94091ad2010201f7418fd2017-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01526-17https://doaj.org/toc/2150-7511ABSTRACT Nearly all virulence factors in Bordetella pertussis are activated by a master two-component system, BvgAS, composed of the sensor kinase BvgS and the response regulator BvgA. When BvgS is active, BvgA is phosphorylated (BvgA~P), and virulence-activated genes (vags) are expressed [Bvg(+) mode]. When BvgS is inactive and BvgA is not phosphorylated, virulence-repressed genes (vrgs) are induced [Bvg(−) mode]. Here, we have used transcriptome sequencing (RNA-seq) and reverse transcription-quantitative PCR (RT-qPCR) to define the BvgAS-dependent regulon of B. pertussis Tohama I. Our analyses reveal more than 550 BvgA-regulated genes, of which 353 are newly identified. BvgA-activated genes include those encoding two-component systems (such as kdpED), multiple other transcriptional regulators, and the extracytoplasmic function (ECF) sigma factor brpL, which is needed for type 3 secretion system (T3SS) expression, further establishing the importance of BvgA~P as an apex regulator of transcriptional networks promoting virulence. Using in vitro transcription, we demonstrate that the promoter for brpL is directly activated by BvgA~P. BvgA-FeBABE cleavage reactions identify BvgA~P binding sites centered at positions −41.5 and −63.5 in bprL. Most importantly, we show for the first time that genes for multiple and varied metabolic pathways are significantly upregulated in the B. pertussis Bvg(−) mode. These include genes for fatty acid and lipid metabolism, sugar and amino acid transporters, pyruvate dehydrogenase, phenylacetic acid degradation, and the glycolate/glyoxylate utilization pathway. Our results suggest that metabolic changes in the Bvg(−) mode may be participating in bacterial survival, transmission, and/or persistence and identify over 200 new vrgs that can be tested for function. IMPORTANCE Within the past 20 years, outbreaks of whooping cough, caused by Bordetella pertussis, have led to respiratory disease and infant mortalities, despite good vaccination coverage. This is due, at least in part, to the introduction of a less effective acellular vaccine in the 1990s. It is crucial, then, to understand the molecular basis of B. pertussis growth and infection. The two-component system BvgA (response regulator)/BvgS (histidine kinase) is the master regulator of B. pertussis virulence genes. We report here the first RNA-seq analysis of the BvgAS regulon in B. pertussis, revealing that more than 550 genes are regulated by BvgAS. We show that genes for multiple and varied metabolic pathways are highly regulated in the Bvg(−) mode (absence of BvgA phosphorylation). Our results suggest that metabolic changes in the Bvg(−) mode may be participating in bacterial survival, transmission, and/or persistence.Kyung MoonRichard P. BonocoraDavid D. KimQing ChenJoseph T. WadeScott StibitzDeborah M. HintonAmerican Society for MicrobiologyarticleBvgAS regulonRNA polymeraseRNA-seqbrpLpertussisMicrobiologyQR1-502ENmBio, Vol 8, Iss 5 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	BvgAS regulon RNA polymerase RNA-seq brpL pertussis Microbiology QR1-502
spellingShingle	BvgAS regulon RNA polymerase RNA-seq brpL pertussis Microbiology QR1-502 Kyung Moon Richard P. Bonocora David D. Kim Qing Chen Joseph T. Wade Scott Stibitz Deborah M. Hinton The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
description	ABSTRACT Nearly all virulence factors in Bordetella pertussis are activated by a master two-component system, BvgAS, composed of the sensor kinase BvgS and the response regulator BvgA. When BvgS is active, BvgA is phosphorylated (BvgA~P), and virulence-activated genes (vags) are expressed [Bvg(+) mode]. When BvgS is inactive and BvgA is not phosphorylated, virulence-repressed genes (vrgs) are induced [Bvg(−) mode]. Here, we have used transcriptome sequencing (RNA-seq) and reverse transcription-quantitative PCR (RT-qPCR) to define the BvgAS-dependent regulon of B. pertussis Tohama I. Our analyses reveal more than 550 BvgA-regulated genes, of which 353 are newly identified. BvgA-activated genes include those encoding two-component systems (such as kdpED), multiple other transcriptional regulators, and the extracytoplasmic function (ECF) sigma factor brpL, which is needed for type 3 secretion system (T3SS) expression, further establishing the importance of BvgA~P as an apex regulator of transcriptional networks promoting virulence. Using in vitro transcription, we demonstrate that the promoter for brpL is directly activated by BvgA~P. BvgA-FeBABE cleavage reactions identify BvgA~P binding sites centered at positions −41.5 and −63.5 in bprL. Most importantly, we show for the first time that genes for multiple and varied metabolic pathways are significantly upregulated in the B. pertussis Bvg(−) mode. These include genes for fatty acid and lipid metabolism, sugar and amino acid transporters, pyruvate dehydrogenase, phenylacetic acid degradation, and the glycolate/glyoxylate utilization pathway. Our results suggest that metabolic changes in the Bvg(−) mode may be participating in bacterial survival, transmission, and/or persistence and identify over 200 new vrgs that can be tested for function. IMPORTANCE Within the past 20 years, outbreaks of whooping cough, caused by Bordetella pertussis, have led to respiratory disease and infant mortalities, despite good vaccination coverage. This is due, at least in part, to the introduction of a less effective acellular vaccine in the 1990s. It is crucial, then, to understand the molecular basis of B. pertussis growth and infection. The two-component system BvgA (response regulator)/BvgS (histidine kinase) is the master regulator of B. pertussis virulence genes. We report here the first RNA-seq analysis of the BvgAS regulon in B. pertussis, revealing that more than 550 genes are regulated by BvgAS. We show that genes for multiple and varied metabolic pathways are highly regulated in the Bvg(−) mode (absence of BvgA phosphorylation). Our results suggest that metabolic changes in the Bvg(−) mode may be participating in bacterial survival, transmission, and/or persistence.
format	article
author	Kyung Moon Richard P. Bonocora David D. Kim Qing Chen Joseph T. Wade Scott Stibitz Deborah M. Hinton
author_facet	Kyung Moon Richard P. Bonocora David D. Kim Qing Chen Joseph T. Wade Scott Stibitz Deborah M. Hinton
author_sort	Kyung Moon
title	The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
title_short	The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
title_full	The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
title_fullStr	The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
title_full_unstemmed	The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>
title_sort	bvgas regulon of <italic toggle="yes">bordetella pertussis</italic>
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/0ad66eba78e94091ad2010201f7418fd
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The BvgAS Regulon of <italic toggle="yes">Bordetella pertussis</italic>

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