Role for the major outer-membrane protein from Vibrio anguillarum in bile resistance and biofilm formation.
Vibrio anguillarum, a fish pathogen, produces a 38 kDa major outer-membrane porin, which may be involved in environmental adaptation. The gene encoding the 38 kDa porin was cloned and deleted. The deduced protein sequence was 75 % identical to that of the major outer-membrane protein (OMP), OmpU, from Vibrio cholerae. LacZ expression from an ompU : : lacZ transcriptional gene fusion was increased 1.5-fold in the presence of bile salts and was decreased 50- to 100-fold in a toxR mutant compared to that in the wild-type, showing that ompU expression is positively regulated by ToxR and induced by bile salts. Similar to a toxR mutant, an ompU mutant showed a slight decrease in motility, an increased sensitivity to bile salts and a thicker biofilm with better surface area coverage compared to that of the wild-type. When ompU was expressed under a ToxR-independent promoter in the toxR mutant, the phenotypes for bile resistance and biofilm formation, but not motility were complemented to that of the wild-type. In rainbow trout, the ompU mutant showed wild-type virulence via immersion into infected seawater and intraperitoneal injection. The ompU mutant produced two colony morphologies: opaque, which did not grow at 0.2 % bile, and translucent, which grew at 2 % bile. The translucent ompU mutant strain produced a second major OMP that was induced by bile. All ompU mutants showed variations in the amount and length of smooth LPS. In V. anguillarum, OmpU is not required for virulence, possibly due to a second OMP also critical for resistance to bile; however, outside of the fish host, OmpU limits the progression of biofilm formation.
Wang
Su-Yan
SY
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
Lauritz
Johan
J
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
Jass
Jana
J
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
Milton
Debra L
DL
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
D018829:Adhesins, Bacterial / Q000378:metabolism; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001646:Bile; D018441:Biofilms / Q000254:growth & development; D024881:Drug Resistance, Bacterial; D005393:Fish Diseases / Q000382:microbiology; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss; D017422:Sequence Analysis, DNA; D014733:Vibrio; D014735:Vibrio Infections / Q000662:veterinary*; D014774:Virulence
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; C096349:OmpU protein, Vibrio cholerae