title: The HtrA-like protease CD3284 modulates virulence of Clostridium difficile.
abstract: In the past decade, Clostridium difficile has emerged as an important gut pathogen. Symptoms of C. difficile infection range from mild diarrhea to pseudomembranous colitis. Besides the two main virulence factors toxin A and toxin B, other virulence factors are likely to play a role in the pathogenesis of the disease. In other Gram-positive and Gram-negative pathogenic bacteria, conserved high-temperature requirement A (HtrA)-like proteases have been shown to have a role in protein homeostasis and quality control. This affects the functionality of virulence factors and the resistance of bacteria to (host-induced) environmental stresses. We found that the C. difficile 630 genome encodes a single HtrA-like protease (CD3284; HtrA) and have analyzed its role in vivo and in vitro through the creation of an isogenic ClosTron-based htrA mutant of C. difficile strain 630Δerm (wild type). In contrast to the attenuated phenotype seen with htrA deletion in other pathogens, this mutant showed enhanced virulence in the Golden Syrian hamster model of acute C. difficile infection. Microarray data analysis showed a pleiotropic effect of htrA on the transcriptome of C. difficile, including upregulation of the toxin A gene. In addition, the htrA mutant showed reduced spore formation and adherence to colonic cells. Together, our data show that htrA can modulate virulence in C. difficile.
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Dennis
authors.initials: D
authors.lastname: Bakker
authors.affiliation: Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Buckley
authors.affiliation: Department of Molecular Genetics, University of Groningen, Groningen, The Netherlands.
authors.forename: Anne
authors.initials: A
authors.lastname: de Jong
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Vincent J C
authors.initials: VJ
authors.lastname: van Winden
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Joost P A
authors.initials: JP
authors.lastname: Verhoeks
authors.affiliation: Department of Molecular Genetics, University of Groningen, Groningen, The Netherlands.
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
authors.affiliation: Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Gillian R
authors.initials: GR
authors.lastname: Douce
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Ed J
authors.initials: EJ
authors.lastname: Kuijper
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands W.K.Smits@LUMC.nl J.corver@LUMC.nl.
authors.forename: Wiep Klaas
authors.initials: WK
authors.lastname: Smits
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, Leiden, The Netherlands W.K.Smits@LUMC.nl J.corver@LUMC.nl.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Corver
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D016360:Clostridioides difficile / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D003015:Clostridium Infections / Q000382:microbiology / Q000473:pathology; D006224:Cricetinae; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000382:microbiology; D005260:Female; D017353:Gene Deletion; D020869:Gene Expression Profiling; D006801:Humans; D008647:Mesocricetus; D046228:Microarray Analysis; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D037521:Virulence Factors; D010447:Peptide Hydrolases