Project description:The bioluminescent bacterium Vibrio fischeri initiates a specific, persistent symbiosis in the light organ of the squid Euprymna scolopes. During the early stages of colonization, V. fischeri is exposed to host-derived nitric oxide (NO). While NO can be both an antimicrobial component of innate immunity and a common signaling molecule of eukaryotes, its roles in beneficial host-microbe associations remain undescribed. V. fischeri encodes HnoX, a member of a family of bacterial NO-binding proteins of unknown function. We hypothesized that HnoX acts as a NO sensor that is involved in regulating symbiosis-related genes during initiation of symbiosis. With an aim to discover the genes whose regulations respond to NO signal, and in an HnoX-mediated fashion in particular, we carried out a whole-genome expression study on the wild-type and an insertional mutant of hnoX.
Project description:The bioluminescent bacterium Vibrio fischeri initiates a specific, persistent symbiosis in the light organ of the squid Euprymna scolopes. During the early stages of colonization, V. fischeri is exposed to host-derived nitric oxide (NO). While NO can be both an antimicrobial component of innate immunity and a common signaling molecule of eukaryotes, its roles in beneficial host-microbe associations remain undescribed. V. fischeri encodes HnoX, a member of a family of bacterial NO-binding proteins of unknown function. We hypothesized that HnoX acts as a NO sensor that is involved in regulating symbiosis-related genes during initiation of symbiosis. With an aim to discover the genes whose regulations respond to NO signal, and in an HnoX-mediated fashion in particular, we carried out a whole-genome expression study on the wild-type and an insertional mutant of hnoX. The wild-type parent and an insertional mutant (hnoX-) of the hnoX gene were grown to early log phase in a minimal-salts medium. One half of each culture was treated with 80µM of the NO-generator, DEA-NONOate, and the other half was left untreated as a control. After 30 min, cells from all the cultures were fixed with RNAprotect Bacteria Reagent. Total RNA was isolated, labeled and hybridized to the Custom Vibrio fischeri GeneChip Array (Affymetrix). Three independent experiments were performed on separate days for statistical analysis.
Project description:The marine bacterium Vibrio fischeri requires flagellar motility to undergo symbiotic initiation with its host, the Hawaiian bobtail squid Euprymna scolopes. We sought to identify the genes activated by the sigma54-dependent flagellar master regulator, FlrA, in V. fischeri, thereby determining the flagellar regulon in this model symbiont. We performed microarray analysis on wild-type Vibrio fischeri ES114 and a flrA deletion mutant, DM159, grown to mid-log phase in seawater tryptone, a condition in which cells are highly motile (two biological replicates per condition).