title: The chitinolytic activity of Listeria monocytogenes EGD is regulated by carbohydrates but also by the virulence regulator PrfA.
abstract: Chitin, an insoluble polymer of N-acetyl-D-glucosamine (GlcNAc), is one of the most abundant carbohydrate polymers in marine and terrestrial environments. Chitin hydrolysis by Listeria monocytogenes depends on two chitinase-encoding genes, chiA and chiB, and the aim of this study was to investigate their regulation. Chitin induces the expression of both chitinases in late exponential growth phase, and chiA but not chiB is furthermore induced by the monomer GlcNAc. Furthermore, their expression is subjected to catabolite control. Chitinases expressed by bacterial pathogens have proven to be important not only for nutrient acquisition and environmental survival but also for infecting animals and humans. Interestingly, the central L. monocytogenes virulence gene regulator, PrfA, is required for the chitinolytic phenotype, as chitinase activity was significantly reduced in prfA mutant cells compared to its level in wild-type cells. In agreement with this, Northern blot analysis showed that the amounts of chiA and chiB transcripts upon induction by chitin were significantly lower in the prfA mutant than in the wild type. The chitinolytic activity and chiA and chiB expression were reduced in the absence of the sigB gene, indicating that σ(B) is also important for the production of chitinases. The chiA, chiB, and chiA chiB mutants were not impaired for in vitro adhesion and invasion in epithelial cell lines, but the chiA chiB double mutant showed less survival ability in a chitin-enriched medium. The regulation of chitinolytic activity in L. monocytogenes is complex, and taken together, the results indicate that the biological role of this activity may not be limited to the external environment.
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark. mhl@life.ku.dk
authors.forename: M H
authors.initials: MH
authors.lastname: Larsen
authors.forename: J J
authors.initials: JJ
authors.lastname: Leisner
authors.forename: H
authors.initials: H
authors.lastname: Ingmer
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000502:physiology*; D015152:Blotting, Northern; D002460:Cell Line; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000096:biosynthesis*; D004847:Epithelial Cells / Q000382:microbiology; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008089:Listeria monocytogenes / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D010454:Peptide Termination Factors / Q000172:deficiency / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D010454:Peptide Termination Factors; C545356:PrfA protein, Listeria monocytogenes; D012333:RNA, Messenger; D002686:Chitin; D002688:Chitinases; D000117:Acetylglucosamine