title: Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence.
abstract: Flavobacterium psychrophilum is an important fish pathogen, responsible for Cold Water Disease, with a significant economic impact on salmonid farms worldwide. In spite of this, little is known about the bacterial physiology and pathogenesis mechanisms, maybe because it is difficult to manipulate, being considered a fastidious microorganism. Mutants obtained using a Tn4351 transposon were screened in order to identify those with alteration in colony morphology, colony spreading and extracellular proteolytic activity, amongst other phenotypes. A F. psychrophilum mutant lacking gliding motility showed interruption of the FP1638 locus that encodes a putative type-2 glycosyltransferase (from here on referred to as fpgA gene, Flavobacterium psychrophilum glycosyltransferase). Additionally, the mutant also showed a decrease in the extracellular proteolytic activity as a consequence of down regulation in the fpgA mutant background of the fpp2-fpp1 operon promoter, responsible for the major extracellular proteolytic activity of the bacterium. The protein glycosylation profile of the parental strain showed the presence of a 22 kDa glycosylated protein which is lost in the mutant. Complementation with the fpgA gene led to the recovery of the wild-type phenotype. LD50 experiments in the rainbow trout infection model show that the mutant was highly attenuated. The pleiotropic phenotype of the mutant demonstrated the importance of this glycosyltranferase in the physiology and virulence of the bacterium. Moreover, the fpgA mutant strain could be considered a good candidate for the design of an attenuated vaccine.
authors.affiliation: Área de Microbiología, Departamento de Biología Funcional, Facultad de Medicina, IUBA, Universidad de Oviedo, Oviedo, 33006, Spain. david.perez@jouy.inra.fr.
authors.forename: David
authors.initials: D
authors.lastname: Pérez-Pascual
authors.forename: Esther
authors.initials: E
authors.lastname: Gómez
authors.forename: José A
authors.initials: JA
authors.lastname: Guijarro
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D005393:Fish Diseases / Q000201:enzymology / Q000382:microbiology*; D045826:Flavobacteriaceae Infections / Q000201:enzymology / Q000382:microbiology / Q000662:veterinary*; D005417:Flavobacterium / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D016695:Glycosyltransferases / Q000235:genetics* / Q000378:metabolism; D007928:Lethal Dose 50; D017686:Oncorhynchus mykiss; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D016695:Glycosyltransferases