The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation

ABSTRACT The spore-forming bacterial pathogen Clostridium difficile is a leading cause of health care-associated infections in the United States. In order for this obligate anaerobe to transmit infection, it must form metabolically dormant spores prior to exiting the host. A key step during this pro...

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Autores principales:	John W. Ribis, Priyanka Ravichandran, Emily E. Putnam, Keyan Pishdadian, Aimee Shen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Clostridium difficile SpoIVA SpoVM coat morphogenesis spore Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/50b6f33f67094b6cbc18745c5330c938
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spelling	oai:doaj.org-article:50b6f33f67094b6cbc18745c5330c9382021-11-15T15:22:05ZThe Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation10.1128/mSphere.00315-172379-5042https://doaj.org/article/50b6f33f67094b6cbc18745c5330c9382017-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00315-17https://doaj.org/toc/2379-5042ABSTRACT The spore-forming bacterial pathogen Clostridium difficile is a leading cause of health care-associated infections in the United States. In order for this obligate anaerobe to transmit infection, it must form metabolically dormant spores prior to exiting the host. A key step during this process is the assembly of a protective, multilayered proteinaceous coat around the spore. Coat assembly depends on coat morphogenetic proteins recruiting distinct subsets of coat proteins to the developing spore. While 10 coat morphogenetic proteins have been identified in Bacillus subtilis, only two of these morphogenetic proteins have homologs in the Clostridia: SpoIVA and SpoVM. C. difficile SpoIVA is critical for proper coat assembly and functional spore formation, but the requirement for SpoVM during this process was unknown. Here, we show that SpoVM is largely dispensable for C. difficile spore formation, in contrast with B. subtilis. Loss of C. difficile SpoVM resulted in modest decreases (~3-fold) in heat- and chloroform-resistant spore formation, while morphological defects such as coat detachment from the forespore and abnormal cortex thickness were observed in ~30% of spoVM mutant cells. Biochemical analyses revealed that C. difficile SpoIVA and SpoVM directly interact, similarly to their B. subtilis counterparts. However, in contrast with B. subtilis, C. difficile SpoVM was not essential for SpoIVA to encase the forespore. Since C. difficile coat morphogenesis requires SpoIVA-interacting protein L (SipL), which is conserved exclusively in the Clostridia, but not the more broadly conserved SpoVM, our results reveal another key difference between C. difficile and B. subtilis spore assembly pathways. IMPORTANCE The spore-forming obligate anaerobe Clostridium difficile is the leading cause of antibiotic-associated diarrheal disease in the United States. When C. difficile spores are ingested by susceptible individuals, they germinate within the gut and transform into vegetative, toxin-secreting cells. During infection, C. difficile must also induce spore formation to survive exit from the host. Since spore formation is essential for transmission, understanding the basic mechanisms underlying sporulation in C. difficile could inform the development of therapeutic strategies targeting spores. In this study, we determine the requirement of the C. difficile homolog of SpoVM, a protein that is essential for spore formation in Bacillus subtilis due to its regulation of coat and cortex formation. We observed that SpoVM plays a minor role in C. difficile spore formation, in contrast with B. subtilis, indicating that this protein would not be a good target for inhibiting spore formation.John W. RibisPriyanka RavichandranEmily E. PutnamKeyan PishdadianAimee ShenAmerican Society for MicrobiologyarticleClostridium difficileSpoIVASpoVMcoatmorphogenesissporeMicrobiologyQR1-502ENmSphere, Vol 2, Iss 5 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Clostridium difficile SpoIVA SpoVM coat morphogenesis spore Microbiology QR1-502
spellingShingle	Clostridium difficile SpoIVA SpoVM coat morphogenesis spore Microbiology QR1-502 John W. Ribis Priyanka Ravichandran Emily E. Putnam Keyan Pishdadian Aimee Shen The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
description	ABSTRACT The spore-forming bacterial pathogen Clostridium difficile is a leading cause of health care-associated infections in the United States. In order for this obligate anaerobe to transmit infection, it must form metabolically dormant spores prior to exiting the host. A key step during this process is the assembly of a protective, multilayered proteinaceous coat around the spore. Coat assembly depends on coat morphogenetic proteins recruiting distinct subsets of coat proteins to the developing spore. While 10 coat morphogenetic proteins have been identified in Bacillus subtilis, only two of these morphogenetic proteins have homologs in the Clostridia: SpoIVA and SpoVM. C. difficile SpoIVA is critical for proper coat assembly and functional spore formation, but the requirement for SpoVM during this process was unknown. Here, we show that SpoVM is largely dispensable for C. difficile spore formation, in contrast with B. subtilis. Loss of C. difficile SpoVM resulted in modest decreases (~3-fold) in heat- and chloroform-resistant spore formation, while morphological defects such as coat detachment from the forespore and abnormal cortex thickness were observed in ~30% of spoVM mutant cells. Biochemical analyses revealed that C. difficile SpoIVA and SpoVM directly interact, similarly to their B. subtilis counterparts. However, in contrast with B. subtilis, C. difficile SpoVM was not essential for SpoIVA to encase the forespore. Since C. difficile coat morphogenesis requires SpoIVA-interacting protein L (SipL), which is conserved exclusively in the Clostridia, but not the more broadly conserved SpoVM, our results reveal another key difference between C. difficile and B. subtilis spore assembly pathways. IMPORTANCE The spore-forming obligate anaerobe Clostridium difficile is the leading cause of antibiotic-associated diarrheal disease in the United States. When C. difficile spores are ingested by susceptible individuals, they germinate within the gut and transform into vegetative, toxin-secreting cells. During infection, C. difficile must also induce spore formation to survive exit from the host. Since spore formation is essential for transmission, understanding the basic mechanisms underlying sporulation in C. difficile could inform the development of therapeutic strategies targeting spores. In this study, we determine the requirement of the C. difficile homolog of SpoVM, a protein that is essential for spore formation in Bacillus subtilis due to its regulation of coat and cortex formation. We observed that SpoVM plays a minor role in C. difficile spore formation, in contrast with B. subtilis, indicating that this protein would not be a good target for inhibiting spore formation.
format	article
author	John W. Ribis Priyanka Ravichandran Emily E. Putnam Keyan Pishdadian Aimee Shen
author_facet	John W. Ribis Priyanka Ravichandran Emily E. Putnam Keyan Pishdadian Aimee Shen
author_sort	John W. Ribis
title	The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
title_short	The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
title_full	The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
title_fullStr	The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
title_full_unstemmed	The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation
title_sort	conserved spore coat protein spovm is largely dispensable in <named-content content-type="genus-species">clostridium difficile</named-content> spore formation
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/50b6f33f67094b6cbc18745c5330c938
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The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in <named-content content-type="genus-species">Clostridium difficile</named-content> Spore Formation

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