Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE

ABSTRACT Most bacteria respond to surfaces by biogenesis of intracellular c-di-GMP, which inhibits motility and induces secretion of biofilm-promoting adherence factors. Bacterial cellulose is a widespread biofilm component whose secretion in Gram-negative species requires an inner membrane, c-di-GM...

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Autores principales:	Samira Zouhir, Wiem Abidi, Meryem Caleechurn, Petya Violinova Krasteva
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	biofilm formation c-di-GMP signaling cellulose secretion structural biology Microbiology QR1-502
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spelling	oai:doaj.org-article:422d496f16084bf281406ffece0a9cca2021-11-15T15:56:44ZStructure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE10.1128/mBio.01303-202150-7511https://doaj.org/article/422d496f16084bf281406ffece0a9cca2020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01303-20https://doaj.org/toc/2150-7511ABSTRACT Most bacteria respond to surfaces by biogenesis of intracellular c-di-GMP, which inhibits motility and induces secretion of biofilm-promoting adherence factors. Bacterial cellulose is a widespread biofilm component whose secretion in Gram-negative species requires an inner membrane, c-di-GMP-dependent synthase tandem (BcsAB), an outer membrane porin (BcsC), and various accessory subunits that regulate synthase assembly and function as well as the exopolysaccharide’s chemical composition and mechanical properties. We recently showed that in Escherichia coli, most Bcs proteins form a megadalton-sized secretory nanomachine, but the role and structure of individual regulatory components remained enigmatic. Here, we demonstrate that essential-for-secretion BcsR and BcsQ regulate each other’s folding and stability and are recruited to the inner membrane via c-di-GMP-sensing BcsE and its intraoperon partner BcsF. Crystallographic and solution-based data show that BcsE’s predicted GIL domain is a degenerate receiver-GGDEF domain tandem (BcsEREC-GGDEF), where the divergent diguanylate cyclase module binds both dimeric c-di-GMP and BcsQ through mutually independent interfaces. In addition, we reveal that a third N-terminal domain (BcsENTD) determines the protein’s homooligomerization and targeting of BcsERQ to the membrane as well as previously unreported interactions with transcription antitermination complex components. Together, the data suggest that BcsE acts on multiple levels to fine-tune bacterial cellulose secretion, from the early stages of secretion system assembly to the maintenance of a membrane-proximal pool of dimeric c-di-GMP for processive synthase activation. IMPORTANCE Bacterial cellulose is a widespread biofilm component that can modulate microbial fitness and virulence both in the environment and infected hosts. Whereas its secretion generally involves an inner membrane c-di-GMP-dependent synthase tandem (BcsAB) across the bacterial domain of life, enterobacteria feature sophisticated Escherichia coli-like Bcs secretion systems, where multiple additional subunits are either required for secretion or contribute to the maximal production of the polysaccharide in vivo. Here, we demonstrate that essential-for-secretion BcsR and BcsQ regulate each other's folding and stability and are recruited to the inner membrane via c-di-GMP-sensing BcsE and its intraoperon partner, BcsF. Crystallographic and functional data reveal that BcsE features unexpected domain architecture and likely acts on multiple levels to fine-tune bacterial cellulose production, from the early stages of secretion system assembly to the maintenence of a membrane-proximal pool of dimeric c-di-GMP for processive synthase activation.Samira ZouhirWiem AbidiMeryem CaleechurnPetya Violinova KrastevaAmerican Society for Microbiologyarticlebiofilm formationc-di-GMP signalingcellulose secretionstructural biologyMicrobiologyQR1-502ENmBio, Vol 11, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	biofilm formation c-di-GMP signaling cellulose secretion structural biology Microbiology QR1-502
spellingShingle	biofilm formation c-di-GMP signaling cellulose secretion structural biology Microbiology QR1-502 Samira Zouhir Wiem Abidi Meryem Caleechurn Petya Violinova Krasteva Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
description	ABSTRACT Most bacteria respond to surfaces by biogenesis of intracellular c-di-GMP, which inhibits motility and induces secretion of biofilm-promoting adherence factors. Bacterial cellulose is a widespread biofilm component whose secretion in Gram-negative species requires an inner membrane, c-di-GMP-dependent synthase tandem (BcsAB), an outer membrane porin (BcsC), and various accessory subunits that regulate synthase assembly and function as well as the exopolysaccharide’s chemical composition and mechanical properties. We recently showed that in Escherichia coli, most Bcs proteins form a megadalton-sized secretory nanomachine, but the role and structure of individual regulatory components remained enigmatic. Here, we demonstrate that essential-for-secretion BcsR and BcsQ regulate each other’s folding and stability and are recruited to the inner membrane via c-di-GMP-sensing BcsE and its intraoperon partner BcsF. Crystallographic and solution-based data show that BcsE’s predicted GIL domain is a degenerate receiver-GGDEF domain tandem (BcsEREC-GGDEF), where the divergent diguanylate cyclase module binds both dimeric c-di-GMP and BcsQ through mutually independent interfaces. In addition, we reveal that a third N-terminal domain (BcsENTD) determines the protein’s homooligomerization and targeting of BcsERQ to the membrane as well as previously unreported interactions with transcription antitermination complex components. Together, the data suggest that BcsE acts on multiple levels to fine-tune bacterial cellulose secretion, from the early stages of secretion system assembly to the maintenance of a membrane-proximal pool of dimeric c-di-GMP for processive synthase activation. IMPORTANCE Bacterial cellulose is a widespread biofilm component that can modulate microbial fitness and virulence both in the environment and infected hosts. Whereas its secretion generally involves an inner membrane c-di-GMP-dependent synthase tandem (BcsAB) across the bacterial domain of life, enterobacteria feature sophisticated Escherichia coli-like Bcs secretion systems, where multiple additional subunits are either required for secretion or contribute to the maximal production of the polysaccharide in vivo. Here, we demonstrate that essential-for-secretion BcsR and BcsQ regulate each other's folding and stability and are recruited to the inner membrane via c-di-GMP-sensing BcsE and its intraoperon partner, BcsF. Crystallographic and functional data reveal that BcsE features unexpected domain architecture and likely acts on multiple levels to fine-tune bacterial cellulose production, from the early stages of secretion system assembly to the maintenence of a membrane-proximal pool of dimeric c-di-GMP for processive synthase activation.
format	article
author	Samira Zouhir Wiem Abidi Meryem Caleechurn Petya Violinova Krasteva
author_facet	Samira Zouhir Wiem Abidi Meryem Caleechurn Petya Violinova Krasteva
author_sort	Samira Zouhir
title	Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
title_short	Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
title_full	Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
title_fullStr	Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
title_full_unstemmed	Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE
title_sort	structure and multitasking of the c-di-gmp-sensing cellulose secretion regulator bcse
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/422d496f16084bf281406ffece0a9cca
work_keys_str_mv	AT samirazouhir structureandmultitaskingofthecdigmpsensingcellulosesecretionregulatorbcse AT wiemabidi structureandmultitaskingofthecdigmpsensingcellulosesecretionregulatorbcse AT meryemcaleechurn structureandmultitaskingofthecdigmpsensingcellulosesecretionregulatorbcse AT petyaviolinovakrasteva structureandmultitaskingofthecdigmpsensingcellulosesecretionregulatorbcse
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Structure and Multitasking of the c-di-GMP-Sensing Cellulose Secretion Regulator BcsE

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