title: Contribution of conserved ATP-dependent proteases of Campylobacter jejuni to stress tolerance and virulence.
abstract: In prokaryotic cells the ATP-dependent proteases Lon and ClpP (Clp proteolytic subunit) are involved in the turnover of misfolded proteins and the degradation of regulatory proteins, and depending on the organism, these proteases contribute variably to stress tolerance. We constructed mutants in the lon and clpP genes of the food-borne human pathogen Campylobacter jejuni and found that the growth of both mutants was impaired at high temperature, a condition known to increase the level of misfolded protein. Moreover, the amounts of misfolded protein aggregates were increased when both proteases were absent, and we propose that both ClpP and Lon are involved in eliminating misfolded proteins in C. jejuni. In order to bind misfolded protein, ClpP has to associate with one of several Clp ATPases. Following inactivation of the ATPase genes clpA and clpX, only the clpX mutant displayed the same heat sensitivity as the clpP mutant, indicating that the ClpXP proteolytic complex is responsible for the degradation of heat-damaged proteins in C. jejuni. Notably, ClpP and ClpX are required for growth at 42 degrees C, which is the temperature of the intestinal tract of poultry, one of the primary carriers of C. jejuni. Thus, ClpP and ClpX may be suitable targets of new intervention strategies aimed at reducing C. jejuni in poultry production. Further characterization of the clpP and lon mutants revealed other altered phenotypes, such as reduced motility, less autoagglutination, and lower levels of invasion of INT407 epithelial cells, suggesting that the proteases may contribute to the virulence of C. jejuni.
authors.affiliation: Department of Veterinary Pathobiology, Faculty of Life Sciences, Stigbøjlen 4, DK-1870 Frederiksberg C, Denmark.
authors.forename: Marianne Thorup
authors.initials: MT
authors.lastname: Cohn
authors.forename: Hanne
authors.initials: H
authors.lastname: Ingmer
authors.forename: Francis
authors.initials: F
authors.lastname: Mulholland
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jørgensen
authors.forename: Jerry M
authors.initials: JM
authors.lastname: Wells
authors.forename: Lone
authors.initials: L
authors.lastname: Brøndsted
mesh_terms: D049069:ATP-Dependent Proteases / Q000235:genetics / Q000502:physiology*; D000222:Adaptation, Physiological; D001422:Bacterial Adhesion / Q000235:genetics / Q000502:physiology; D001426:Bacterial Proteins / Q000378:metabolism; D016123:Campylobacter jejuni / Q000472:pathogenicity* / Q000502:physiology*; D002460:Cell Line; D049071:Endopeptidase Clp / Q000235:genetics / Q000502:physiology; D004847:Epithelial Cells / Q000382:microbiology; D017353:Gene Deletion; D006358:Hot Temperature; D006801:Humans; D008124:Locomotion / Q000235:genetics / Q000502:physiology; D010447:Peptide Hydrolases / Q000235:genetics / Q000502:physiology; D049070:Protease La / Q000235:genetics / Q000502:physiology; D017510:Protein Folding; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; C526033:ClpA protein, Campylobacter jejuni; D010447:Peptide Hydrolases; D049069:ATP-Dependent Proteases; D049070:Protease La; D049071:Endopeptidase Clp; C526034:ClpX protein, Campylobacter jejuni