Inducing and Quantifying Clostridium difficile Spore Formation

2016 ◽  
pp. 129-142 ◽  
Author(s):  
Aimee Shen ◽  
Kelly A. Fimlaid ◽  
Keyan Pishdadian
Keyword(s):  
Clostridium Difficile ◽  
Spore Formation

scholarly journals Genome-Wide Analysis of Cell Type-Specific Gene Transcription during Spore Formation in Clostridium difficile

PLoS Genetics ◽  
2013 ◽  
Vol 9 (10) ◽  
pp. e1003756 ◽  
Author(s):  
Laure Saujet ◽  
Fátima C. Pereira ◽  
Monica Serrano ◽  
Olga Soutourina ◽  
Marc Monot ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Gene Transcription ◽  
Spore Formation ◽  
Specific Gene ◽  
Cell Type ◽  
Genome Wide Analysis ◽  
Genome Wide ◽  
Cell Type Specific

scholarly journals Regulation of Clostridium difficile Spore Formation by the SpoIIQ and SpoIIIA Proteins

PLoS Genetics ◽  
2015 ◽  
Vol 11 (10) ◽  
pp. e1005562 ◽  
Author(s):  
Kelly A. Fimlaid ◽  
Owen Jensen ◽  
M. Lauren Donnelly ◽  
M. Sloan Siegrist ◽  
Aimee Shen
Keyword(s):  
Clostridium Difficile ◽  
Spore Formation

Hospital disinfectants and spore formation by Clostridium difficile

The Lancet ◽  
2000 ◽  
Vol 356 (9238) ◽  
pp. 1324 ◽  
Author(s):  
Mark H Wilcox ◽  
Warren N Fawley
Keyword(s):  
Clostridium Difficile ◽  
Spore Formation

scholarly journals Spore Formation and Toxin Production in Clostridium difficile Biofilms

PLoS ONE ◽  
2014 ◽  
Vol 9 (1) ◽  
pp. e87757 ◽  
Author(s):  
Ekaterina G. Semenyuk ◽  
Michelle L. Laning ◽  
Jennifer Foley ◽  
Pehga F. Johnston ◽  
Katherine L. Knight ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Toxin Production ◽  
Spore Formation

scholarly journals The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in Clostridium difficile Spore Formation

mSphere ◽  
2017 ◽  
Vol 2 (5) ◽  
Author(s):  
John W. Ribis ◽  
Priyanka Ravichandran ◽  
Emily E. Putnam ◽  
Keyan Pishdadian ◽  
Aimee Shen
Keyword(s):  
United States ◽  
Bacillus Subtilis ◽  
Clostridium Difficile ◽  
Diarrheal Disease ◽  
The United States ◽  
Spore Formation ◽  
Minor Role ◽  
Obligate Anaerobe ◽  
Content Type ◽  
A Minor

ABSTRACT 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. 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.

scholarly journals Characterization of Clostridium difficile Spores Lacking Either SpoVAC or Dipicolinic Acid Synthetase

2016 ◽  
Vol 198 (11) ◽  
pp. 1694-1707 ◽  
Author(s):  
M. Lauren Donnelly ◽  
Kelly A. Fimlaid ◽  
Aimee Shen
Keyword(s):  
Clostridium Difficile ◽  
Heat Resistance ◽  
Spore Germination ◽  
Dipicolinic Acid ◽  
The United States ◽  
Spore Formation ◽  
Obligate Anaerobe ◽  
Wild Type ◽  
Content Type ◽  
Wet Heat

ABSTRACTThe spore-forming obligate anaerobeClostridium difficileis a leading cause of antibiotic-associated diarrhea around the world. In order forC. difficileto cause infection, its metabolically dormant spores must germinate in the gastrointestinal tract. During germination, spores degrade their protective cortex peptidoglycan layers, release dipicolinic acid (DPA), and hydrate their cores. InC. difficile, cortex hydrolysis is necessary for DPA release, whereas inBacillus subtilis, DPA release is necessary for cortex hydrolysis. Given this difference, we tested whether DPA synthesis and/or release was required forC. difficilespore germination by constructing mutations in eitherspoVACordpaAB, which encode an ion channel predicted to transport DPA into the forespore and the enzyme complex predicted to synthesize DPA, respectively.C. difficilespoVACanddpaABmutant spores lacked DPA but could be stably purified and were more hydrated than wild-type spores; in contrast,B. subtilisspoVACanddpaABmutant spores were unstable. AlthoughC. difficilespoVACanddpaABmutant spores exhibited wild-type germination responses, they were more readily killed by wet heat. Cortex hydrolysis was not affected by this treatment, indicating that wet heat inhibits a stage downstream of this event. Interestingly,C. difficilespoVACmutant spores were significantly more sensitive to heat treatment thandpaABmutant spores, indicating that SpoVAC plays additional roles in conferring heat resistance. Taken together, our results demonstrate that SpoVAC and DPA synthetase controlC. difficilespore resistance and reveal differential requirements for these proteins among theFirmicutes.IMPORTANCEClostridium difficileis a spore-forming obligate anaerobe that causes ∼500,000 infections per year in the United States. Although spore germination is essential forC. difficileto cause disease, the factors required for this process have been only partially characterized. This study describes the roles of two factors, DpaAB and SpoVAC, which control the synthesis and release of dipicolinic acid (DPA), respectively, from bacterial spores. Previous studies of these proteins in other spore-forming organisms indicated that they are differentially required for spore formation, germination, and resistance. We now show that the proteins are dispensable forC. difficilespore formation and germination but are necessary for heat resistance. Thus, our study further highlights the diverse functions of DpaAB and SpoVAC in spore-forming organisms.

Clostridium difficile Infections May Be Increasing

2010 ◽  
Vol 44 (4) ◽  
pp. 13
Author(s):  
SHARON WORCESTER
Keyword(s):  
Clostridium Difficile
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