Structural and functional analysis of the CspB protease required for Clostridium spore germination.

Spores are the major transmissive form of the nosocomial pathogen Clostridium difficile, a leading cause of healthcare-associated diarrhea worldwide. Successful transmission of C. difficile requires that its hardy, resistant spores germinate into vegetative cells in the gastrointestinal tract. A cri...

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Autores principales:	Chloe M Adams, Brian E Eckenroth, Emily E Putnam, Sylvie Doublié, Aimee Shen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/ab18d43086654511981358cfea09d71f
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spelling	oai:doaj.org-article:ab18d43086654511981358cfea09d71f2021-11-18T06:06:03ZStructural and functional analysis of the CspB protease required for Clostridium spore germination.1553-73661553-737410.1371/journal.ppat.1003165https://doaj.org/article/ab18d43086654511981358cfea09d71f2013-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23408892/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Spores are the major transmissive form of the nosocomial pathogen Clostridium difficile, a leading cause of healthcare-associated diarrhea worldwide. Successful transmission of C. difficile requires that its hardy, resistant spores germinate into vegetative cells in the gastrointestinal tract. A critical step during this process is the degradation of the spore cortex, a thick layer of peptidoglycan surrounding the spore core. In Clostridium sp., cortex degradation depends on the proteolytic activation of the cortex hydrolase, SleC. Previous studies have implicated Csps as being necessary for SleC cleavage during germination; however, their mechanism of action has remained poorly characterized. In this study, we demonstrate that CspB is a subtilisin-like serine protease whose activity is essential for efficient SleC cleavage and C. difficile spore germination. By solving the first crystal structure of a Csp family member, CspB, to 1.6 Å, we identify key structural domains within CspB. In contrast with all previously solved structures of prokaryotic subtilases, the CspB prodomain remains tightly bound to the wildtype subtilase domain and sterically occludes a catalytically competent active site. The structure, combined with biochemical and genetic analyses, reveals that Csp proteases contain a unique jellyroll domain insertion critical for stabilizing the protease in vitro and in C. difficile. Collectively, our study provides the first molecular insight into CspB activity and function. These studies may inform the development of inhibitors that can prevent clostridial spore germination and thus disease transmission.Chloe M AdamsBrian E EckenrothEmily E PutnamSylvie DoubliéAimee ShenPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 9, Iss 2, p e1003165 (2013)
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 Chloe M Adams Brian E Eckenroth Emily E Putnam Sylvie Doublié Aimee Shen Structural and functional analysis of the CspB protease required for Clostridium spore germination.
description	Spores are the major transmissive form of the nosocomial pathogen Clostridium difficile, a leading cause of healthcare-associated diarrhea worldwide. Successful transmission of C. difficile requires that its hardy, resistant spores germinate into vegetative cells in the gastrointestinal tract. A critical step during this process is the degradation of the spore cortex, a thick layer of peptidoglycan surrounding the spore core. In Clostridium sp., cortex degradation depends on the proteolytic activation of the cortex hydrolase, SleC. Previous studies have implicated Csps as being necessary for SleC cleavage during germination; however, their mechanism of action has remained poorly characterized. In this study, we demonstrate that CspB is a subtilisin-like serine protease whose activity is essential for efficient SleC cleavage and C. difficile spore germination. By solving the first crystal structure of a Csp family member, CspB, to 1.6 Å, we identify key structural domains within CspB. In contrast with all previously solved structures of prokaryotic subtilases, the CspB prodomain remains tightly bound to the wildtype subtilase domain and sterically occludes a catalytically competent active site. The structure, combined with biochemical and genetic analyses, reveals that Csp proteases contain a unique jellyroll domain insertion critical for stabilizing the protease in vitro and in C. difficile. Collectively, our study provides the first molecular insight into CspB activity and function. These studies may inform the development of inhibitors that can prevent clostridial spore germination and thus disease transmission.
format	article
author	Chloe M Adams Brian E Eckenroth Emily E Putnam Sylvie Doublié Aimee Shen
author_facet	Chloe M Adams Brian E Eckenroth Emily E Putnam Sylvie Doublié Aimee Shen
author_sort	Chloe M Adams
title	Structural and functional analysis of the CspB protease required for Clostridium spore germination.
title_short	Structural and functional analysis of the CspB protease required for Clostridium spore germination.
title_full	Structural and functional analysis of the CspB protease required for Clostridium spore germination.
title_fullStr	Structural and functional analysis of the CspB protease required for Clostridium spore germination.
title_full_unstemmed	Structural and functional analysis of the CspB protease required for Clostridium spore germination.
title_sort	structural and functional analysis of the cspb protease required for clostridium spore germination.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/ab18d43086654511981358cfea09d71f
work_keys_str_mv	AT chloemadams structuralandfunctionalanalysisofthecspbproteaserequiredforclostridiumsporegermination AT brianeeckenroth structuralandfunctionalanalysisofthecspbproteaserequiredforclostridiumsporegermination AT emilyeputnam structuralandfunctionalanalysisofthecspbproteaserequiredforclostridiumsporegermination AT sylviedoublie structuralandfunctionalanalysisofthecspbproteaserequiredforclostridiumsporegermination AT aimeeshen structuralandfunctionalanalysisofthecspbproteaserequiredforclostridiumsporegermination
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Structural and functional analysis of the CspB protease required for Clostridium spore germination.

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