Identification and characterization of a hemolysin gene cluster in Vibrio anguillarum.

Infection and Immunity
Jessica L Rock, David R Nelson

Abstract

Vibrio anguillarum is a causative agent of vibriosis in fish. Hemolytic activity has been suggested as a virulence factor by contributing to hemorrhagic septicemia and diarrhea. In order to identify and characterize the hemolysin genes and examine the role of hemolytic activity in virulence, a mini-Tn10Kan mutagenesis clone bank of V. anguillarum was screened. While no hemolysin-negative strains were observed, several mutants with two- to threefold-increased hemolytic activity were found. The region containing the insertion mutation was cloned, sequenced, and found to contain the V. anguillarum hemolysin (vah1) and two other open reading frames, coding for a putative lactonizing lipase (llpA) and a putative phospholipase (plp). The mini-Tn10Kan was inserted into plp. Site-directed mutagenesis of each gene revealed that mutations in vah1 and llpA did not affect hemolytic activity, but insertions into plp caused a two- to threefold increase in hemolysis. Double mutations in plp and either vah1 or llpA resulted in wild-type hemolytic activity. Complementation of plp restored hemolytic activity to wild-type levels. Spectrophotometric determination of hemolysin specific activity revealed that activity on a per cell basis peaked duri...Continue Reading

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Citations

Jul 19, 2011·Journal of Bacteriology·Ling LiDavid R Nelson
Jun 1, 2011·Fish Pathology·Hiroaki Naka, Jorge H Crosa
Jan 9, 2007·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Manuel L Lemos, Carlos R Osorio
Feb 22, 2011·Microbial Pathogenesis·Hege Smith TunsjøTrine M L'abée-Lund
Jun 23, 2009·Microbial Pathogenesis·Hege Smith TunsjøTrine Marie L'Abée-Lund
Dec 5, 2015·Genome Announcements·Edward SpinardDavid R Nelson
Aug 16, 2011·Journal of Fish Diseases·I FransH Rediers
Apr 9, 2016·Genome Announcements·Javier DubertJuan L Barja
Jul 30, 2016·Genome Announcements·Edward J SpinardJuan L Barja
Aug 10, 2017·Frontiers in Cellular and Infection Microbiology·Yingjie Li, Qingjun Ma
Dec 22, 2020·PeerJ·Mie Johanne HansenInger Dalsgaard

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