Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm

ABSTRACT Glucose-specific enzyme IIA (EIIAGlc) is a central regulator of bacterial metabolism and an intermediate in the phosphoenolpyruvate phosphotransferase system (PTS), a conserved phosphotransfer cascade that controls carbohydrate transport. We previously reported that EIIAGlc activates transc...

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Autores principales:	Bradley S. Pickering, Daniel R. Smith, Paula I. Watnick
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/117ef15201df45bcbb78e172da75fda7
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spelling	oai:doaj.org-article:117ef15201df45bcbb78e172da75fda72021-11-15T15:39:11ZGlucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm10.1128/mBio.00228-122150-7511https://doaj.org/article/117ef15201df45bcbb78e172da75fda72012-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00228-12https://doaj.org/toc/2150-7511ABSTRACT Glucose-specific enzyme IIA (EIIAGlc) is a central regulator of bacterial metabolism and an intermediate in the phosphoenolpyruvate phosphotransferase system (PTS), a conserved phosphotransfer cascade that controls carbohydrate transport. We previously reported that EIIAGlc activates transcription of the genes required for Vibrio cholerae biofilm formation. While EIIAGlc modulates the function of many proteins through a direct interaction, none of the known regulatory binding partners of EIIAGlc activates biofilm formation. Therefore, we used tandem affinity purification (TAP) to compare binding partners of EIIAGlc in both planktonic and biofilm cells. A surprising number of novel EIIAGlc binding partners were identified predominantly under one condition or the other. Studies of planktonic cells revealed established partners of EIIAGlc, such as adenylate cyclase and glycerol kinase. In biofilms, MshH, a homolog of Escherichia coli CsrD, was found to be a dominant binding partner of EIIAGlc. Further studies revealed that MshH inhibits biofilm formation. This function was independent of the Carbon storage regulator (Csr) pathway and dependent on EIIAGlc. To explore the existence of multiprotein complexes centered on EIIAGlc, we also affinity purified the binding partners of adenylate cyclase from biofilm cells. In addition to EIIAGlc, this analysis yielded many of the same proteins that copurified with EIIAGlc. We hypothesize that EIIAGlc serves as a hub for multiprotein complexes and furthermore that these complexes may provide a mechanism for competitive and cooperative interactions between binding partners. IMPORTANCE EIIAGlc is a global regulator of microbial physiology that acts through direct interactions with other proteins. This work represents the first demonstration that the protein partners of EIIAGlc are distinct in the microbial biofilm. Furthermore, it provides the first evidence that EIIAGlc may exist in multiprotein complexes with its partners, setting the stage for an investigation of how the multiple partners of EIIAGlc influence one another. Last, it provides a connection between the phosphoenolpyruvate phosphotransferase (PTS) and Csr (Carbon storage regulator) regulatory systems. This work increases our understanding of the complexity of regulation by EIIAGlc and provides a link between the PTS and Csr networks, two global regulatory cascades that influence microbial physiology.Bradley S. PickeringDaniel R. SmithPaula I. WatnickAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 6 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Bradley S. Pickering Daniel R. Smith Paula I. Watnick Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
description	ABSTRACT Glucose-specific enzyme IIA (EIIAGlc) is a central regulator of bacterial metabolism and an intermediate in the phosphoenolpyruvate phosphotransferase system (PTS), a conserved phosphotransfer cascade that controls carbohydrate transport. We previously reported that EIIAGlc activates transcription of the genes required for Vibrio cholerae biofilm formation. While EIIAGlc modulates the function of many proteins through a direct interaction, none of the known regulatory binding partners of EIIAGlc activates biofilm formation. Therefore, we used tandem affinity purification (TAP) to compare binding partners of EIIAGlc in both planktonic and biofilm cells. A surprising number of novel EIIAGlc binding partners were identified predominantly under one condition or the other. Studies of planktonic cells revealed established partners of EIIAGlc, such as adenylate cyclase and glycerol kinase. In biofilms, MshH, a homolog of Escherichia coli CsrD, was found to be a dominant binding partner of EIIAGlc. Further studies revealed that MshH inhibits biofilm formation. This function was independent of the Carbon storage regulator (Csr) pathway and dependent on EIIAGlc. To explore the existence of multiprotein complexes centered on EIIAGlc, we also affinity purified the binding partners of adenylate cyclase from biofilm cells. In addition to EIIAGlc, this analysis yielded many of the same proteins that copurified with EIIAGlc. We hypothesize that EIIAGlc serves as a hub for multiprotein complexes and furthermore that these complexes may provide a mechanism for competitive and cooperative interactions between binding partners. IMPORTANCE EIIAGlc is a global regulator of microbial physiology that acts through direct interactions with other proteins. This work represents the first demonstration that the protein partners of EIIAGlc are distinct in the microbial biofilm. Furthermore, it provides the first evidence that EIIAGlc may exist in multiprotein complexes with its partners, setting the stage for an investigation of how the multiple partners of EIIAGlc influence one another. Last, it provides a connection between the phosphoenolpyruvate phosphotransferase (PTS) and Csr (Carbon storage regulator) regulatory systems. This work increases our understanding of the complexity of regulation by EIIAGlc and provides a link between the PTS and Csr networks, two global regulatory cascades that influence microbial physiology.
format	article
author	Bradley S. Pickering Daniel R. Smith Paula I. Watnick
author_facet	Bradley S. Pickering Daniel R. Smith Paula I. Watnick
author_sort	Bradley S. Pickering
title	Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
title_short	Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
title_full	Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
title_fullStr	Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
title_full_unstemmed	Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm
title_sort	glucose-specific enzyme iia has unique binding partners in the <named-content content-type="genus-species">vibrio cholerae</named-content> biofilm
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/117ef15201df45bcbb78e172da75fda7
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Glucose-Specific Enzyme IIA Has Unique Binding Partners in The <named-content content-type="genus-species">Vibrio cholerae</named-content> Biofilm

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