MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.

Klebsiella pneumoniae causes significant morbidity and mortality worldwide, particularly amongst hospitalized individuals. The principle mechanism for pathogenesis in hospital environments involves the formation of biofilms, primarily on implanted medical devices. In this study, we constructed a tra...

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Autores principales:	Jonathan J Wilksch, Ji Yang, Abigail Clements, Jacinta L Gabbe, Kirsty R Short, Hanwei Cao, Rosalia Cavaliere, Catherine E James, Cynthia B Whitchurch, Mark A Schembri, Mary L C Chuah, Zhao-Xun Liang, Odilia L Wijburg, Adam W Jenney, Trevor Lithgow, Richard A Strugnell
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/1b9418132280410ca0559776e41d2207
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spelling	oai:doaj.org-article:1b9418132280410ca0559776e41d22072021-11-18T06:03:05ZMrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.1553-73661553-737410.1371/journal.ppat.1002204https://doaj.org/article/1b9418132280410ca0559776e41d22072011-08-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21901098/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Klebsiella pneumoniae causes significant morbidity and mortality worldwide, particularly amongst hospitalized individuals. The principle mechanism for pathogenesis in hospital environments involves the formation of biofilms, primarily on implanted medical devices. In this study, we constructed a transposon mutant library in a clinical isolate, K. pneumoniae AJ218, to identify the genes and pathways implicated in biofilm formation. Three mutants severely defective in biofilm formation contained insertions within the mrkABCDF genes encoding the main structural subunit and assembly machinery for type 3 fimbriae. Two other mutants carried insertions within the yfiN and mrkJ genes, which encode GGDEF domain- and EAL domain-containing c-di-GMP turnover enzymes, respectively. The remaining two isolates contained insertions that inactivated the mrkH and mrkI genes, which encode for novel proteins with a c-di-GMP-binding PilZ domain and a LuxR-type transcriptional regulator, respectively. Biochemical and functional assays indicated that the effects of these factors on biofilm formation accompany concomitant changes in type 3 fimbriae expression. We mapped the transcriptional start site of mrkA, demonstrated that MrkH directly activates transcription of the mrkA promoter and showed that MrkH binds strongly to the mrkA regulatory region only in the presence of c-di-GMP. Furthermore, a point mutation in the putative c-di-GMP-binding domain of MrkH completely abolished its function as a transcriptional activator. In vivo analysis of the yfiN and mrkJ genes strongly indicated their c-di-GMP-specific function as diguanylate cyclase and phosphodiesterase, respectively. In addition, in vitro assays showed that purified MrkJ protein has strong c-di-GMP phosphodiesterase activity. These results demonstrate for the first time that c-di-GMP can function as an effector to stimulate the activity of a transcriptional activator, and explain how type 3 fimbriae expression is coordinated with other gene expression programs in K. pneumoniae to promote biofilm formation to implanted medical devices.Jonathan J WilkschJi YangAbigail ClementsJacinta L GabbeKirsty R ShortHanwei CaoRosalia CavaliereCatherine E JamesCynthia B WhitchurchMark A SchembriMary L C ChuahZhao-Xun LiangOdilia L WijburgAdam W JenneyTrevor LithgowRichard A StrugnellPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 7, Iss 8, p e1002204 (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 Jonathan J Wilksch Ji Yang Abigail Clements Jacinta L Gabbe Kirsty R Short Hanwei Cao Rosalia Cavaliere Catherine E James Cynthia B Whitchurch Mark A Schembri Mary L C Chuah Zhao-Xun Liang Odilia L Wijburg Adam W Jenney Trevor Lithgow Richard A Strugnell MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
description	Klebsiella pneumoniae causes significant morbidity and mortality worldwide, particularly amongst hospitalized individuals. The principle mechanism for pathogenesis in hospital environments involves the formation of biofilms, primarily on implanted medical devices. In this study, we constructed a transposon mutant library in a clinical isolate, K. pneumoniae AJ218, to identify the genes and pathways implicated in biofilm formation. Three mutants severely defective in biofilm formation contained insertions within the mrkABCDF genes encoding the main structural subunit and assembly machinery for type 3 fimbriae. Two other mutants carried insertions within the yfiN and mrkJ genes, which encode GGDEF domain- and EAL domain-containing c-di-GMP turnover enzymes, respectively. The remaining two isolates contained insertions that inactivated the mrkH and mrkI genes, which encode for novel proteins with a c-di-GMP-binding PilZ domain and a LuxR-type transcriptional regulator, respectively. Biochemical and functional assays indicated that the effects of these factors on biofilm formation accompany concomitant changes in type 3 fimbriae expression. We mapped the transcriptional start site of mrkA, demonstrated that MrkH directly activates transcription of the mrkA promoter and showed that MrkH binds strongly to the mrkA regulatory region only in the presence of c-di-GMP. Furthermore, a point mutation in the putative c-di-GMP-binding domain of MrkH completely abolished its function as a transcriptional activator. In vivo analysis of the yfiN and mrkJ genes strongly indicated their c-di-GMP-specific function as diguanylate cyclase and phosphodiesterase, respectively. In addition, in vitro assays showed that purified MrkJ protein has strong c-di-GMP phosphodiesterase activity. These results demonstrate for the first time that c-di-GMP can function as an effector to stimulate the activity of a transcriptional activator, and explain how type 3 fimbriae expression is coordinated with other gene expression programs in K. pneumoniae to promote biofilm formation to implanted medical devices.
format	article
author	Jonathan J Wilksch Ji Yang Abigail Clements Jacinta L Gabbe Kirsty R Short Hanwei Cao Rosalia Cavaliere Catherine E James Cynthia B Whitchurch Mark A Schembri Mary L C Chuah Zhao-Xun Liang Odilia L Wijburg Adam W Jenney Trevor Lithgow Richard A Strugnell
author_facet	Jonathan J Wilksch Ji Yang Abigail Clements Jacinta L Gabbe Kirsty R Short Hanwei Cao Rosalia Cavaliere Catherine E James Cynthia B Whitchurch Mark A Schembri Mary L C Chuah Zhao-Xun Liang Odilia L Wijburg Adam W Jenney Trevor Lithgow Richard A Strugnell
author_sort	Jonathan J Wilksch
title	MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
title_short	MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
title_full	MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
title_fullStr	MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
title_full_unstemmed	MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
title_sort	mrkh, a novel c-di-gmp-dependent transcriptional activator, controls klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/1b9418132280410ca0559776e41d2207
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MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.

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