title: Reexamining the Germination Phenotypes of Several Clostridium difficile Strains Suggests Another Role for the CspC Germinant Receptor.
abstract: Clostridium difficile spore germination is essential for colonization and disease. The signals that initiate C. difficile spore germination are a combination of taurocholic acid (a bile acid) and glycine. Interestingly, the chenodeoxycholic acid class (CDCA) bile acids competitively inhibit taurocholic acid-mediated germination, suggesting that compounds that inhibit spore germination could be developed into drugs that prophylactically prevent C. difficile infection or reduce recurring disease. However, a recent report called into question the utility of such a strategy to prevent infection by describing C. difficile strains that germinated in the apparent absence of bile acids or germinated in the presence of the CDCA inhibitor. Because the mechanisms of C. difficile spore germination are beginning to be elucidated, the mechanism of germination in these particular strains could yield important information on how C. difficile spores initiate germination. Therefore, we quantified the interaction of these strains with taurocholic acid and CDCA, the rates of spore germination, the release of DPA from the spore core, and the abundance of the germinant receptor complex (CspC, CspB, and SleC). We found that strains previously observed to germinate in the absence of taurocholic acid correspond to more potent 50% effective concentrations (EC50 values; the concentrations that achieve a half-maximum germination rate) of the germinant and are still inhibited by CDCA, possibly explaining the previous observations. By comparing the germination kinetics and the abundance of proteins in the germinant receptor complex, we revised our original model for CspC-mediated activation of spore germination and propose that CspC may activate spore germination and then inhibit downstream processes.
  
  
  OBJECTIVE
  Clostridium difficile forms metabolically dormant spores that persist in the health care environment. In susceptible hosts, C. difficile spores germinate in response to certain bile acids and glycine. Blocking germination by C. difficile spores is an attractive strategy to prevent the initiation of disease or to block recurring infection. However, certain C. difficile strains have been identified whose spores germinate in the absence of bile acids or are not blocked by known inhibitors of C. difficile spore germination (calling into question the utility of such strategies). Here, we further investigate these strains and reestablish that bile acid activators and inhibitors of germination affect these strains and use these data to suggest another role for the C. difficile bile acid germinant receptor.
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Disha
authors.initials: D
authors.lastname: Bhattacharjee
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Francis
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Xicheng
authors.initials: X
authors.lastname: Ding
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Kathleen N
authors.initials: KN
authors.lastname: McAllister
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Ritu
authors.initials: R
authors.lastname: Shrestha
authors.affiliation: Department of Biology, Texas A&M University, College Station, Texas, USA jsorg@bio.tamu.edu.
authors.forename: Joseph A
authors.identifier: http://orcid.org/0000-0001-7822-2656
authors.initials: JA
authors.lastname: Sorg
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002635:Chenodeoxycholic Acid / Q000494:pharmacology*; D002756:Cholagogues and Choleretics / Q000494:pharmacology*; D016360:Clostridioides difficile / Q000145:classification / Q000235:genetics / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D010641:Phenotype; D013171:Spores, Bacterial / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D013656:Taurocholic Acid / Q000494:pharmacology*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D002756:Cholagogues and Choleretics; C102917:CspC protein, Clostridium; D002635:Chenodeoxycholic Acid; D013656:Taurocholic Acid