title: The novel polysaccharide deacetylase homologue Pdi contributes to virulence of the aquatic pathogen Streptococcus iniae.
abstract: The aquatic zoonotic pathogen Streptococcus iniae represents a threat to the worldwide aquaculture industry and poses a risk to humans who handle raw fish. Because little is known about the mechanisms of S. iniae pathogenesis or virulence factors, we established a high-throughput system combining whole-genome pyrosequencing and transposon mutagenesis that allowed us to identify virulence proteins, including Pdi, the polysaccharide deacetylase of S. iniae, that we describe here. Using bioinformatics tools, we identified a highly conserved signature motif in Pdi that is also conserved in the peptidoglycan deacetylase PgdA protein family. A Deltapdi mutant was attenuated for virulence in the hybrid striped bass model and for survival in whole fish blood. Moreover, Pdi was found to promote bacterial resistance to lysozyme killing and the ability to adhere to and invade epithelial cells. On the other hand, there was no difference in the autolytic potential, resistance to oxidative killing or resistance to cationic antimicrobial peptides between S. iniae wild-type and Deltapdi. In conclusion, we have demonstrated that pdi is involved in S. iniae adherence and invasion, lysozyme resistance and survival in fish blood, and have shown that pdi plays a role in the pathogenesis of S. iniae. Identification of Pdi and other S. iniae virulence proteins is a necessary initial step towards the development of appropriate preventive and therapeutic measures against diseases and economic losses caused by this pathogen.
authors.affiliation: Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0687, La Jolla, CA 92093-0687, USA.
authors.forename: Carlo J E
authors.initials: CJE
authors.lastname: Milani
authors.affiliation: Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, 11562 Cairo, Egypt.
authors.forename: Ramy K
authors.initials: RK
authors.lastname: Aziz
authors.affiliation: Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0687, La Jolla, CA 92093-0687, USA.
authors.forename: Jeffrey B
authors.initials: JB
authors.lastname: Locke
authors.affiliation: Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0687, La Jolla, CA 92093-0687, USA.
authors.forename: Samira
authors.initials: S
authors.lastname: Dahesh
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, MC 0657, La Jolla, CA 92093-0657, USA.|Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0687, La Jolla, CA 92093-0687, USA.
authors.forename: Victor
authors.initials: V
authors.lastname: Nizet
authors.affiliation: Aqua Bounty Technologies, 8395 Camino Santa Fe, Suite E, San Diego, CA 92121, USA.|Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0687, La Jolla, CA 92093-0687, USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Buchanan
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001433:Bacteriolysis; D001492:Bass / Q000097:blood / Q000382:microbiology; D001770:Blood Bactericidal Activity; D002460:Cell Line; D004251:DNA Transposable Elements; D004847:Epithelial Cells / Q000382:microbiology; D005393:Fish Diseases / Q000382:microbiology; D018390:Gene Targeting; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism; D016296:Mutagenesis; D017382:Reactive Oxygen Species / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013290:Streptococcal Infections / Q000382:microbiology / Q000662:veterinary; D013291:Streptococcus / Q000201:enzymology / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D017382:Reactive Oxygen Species; D037521:Virulence Factors; D009113:Muramidase; D000581:Amidohydrolases; C022427:polysaccharide deacetylase