Abstract
Vibrio parahaemolyticus is an emerging bacterial pathogen capable of causing inflammatory gastroenteritis, wound infections, and septicemia. As a food-borne illness, infection is most frequently associated with the consumption of raw or undercooked seafood, particularly shellfish. It is the primary cause of Vibrio-associated food-borne illness in the United States and the leading cause of food-borne illness in Japan. The larger of its two chromosomes harbors a set of genes encoding type III section system 1 (T3SS1), a virulence factor present in all V. parahaemolyticus strains that is similar to the Yersinia ysc T3SS. T3SS1 translocates effector proteins into eukaryotic cells where they induce changes to cellular physiology and modulate host-pathogen interactions. T3SS1 is also responsible for cytotoxicity toward several different cultured cell lines as well as mortality in a mouse model. Herein we used RNA-seq to obtain global transcriptome patterns of V. parahaemolyticus under conditions that either induce [growth in Dulbecco's Modified Eagle Medium (DMEM) media, in trans expression of transcriptional regulator exsA] or repress T3SS1 expression (growth in LB-S media, in trans exsD expression) and during infection of HeLa cells over time. Comparative transcriptomic analysis demonstrated notable differences in the expression patterns under inducing conditions and was also used to generate an expression profile of V. parahaemolyticus during infection of HeLa cells. In addition, we identified several new genes that are associated with T3SS1 expression and may warrant further study.
Highlights
Vibrio parahaemolyticus (V. parahaemolyticus) is an emerging food-borne bacterial pathogen that has caused significant disease in several countries
The T3SS is a organelle possessed by several species of Gram-negative bacteria that mediates the transport of specialized proteins, termed “effectors,” directly from the bacterial cytoplasm into the cytosol of eukaryotic cells (Galán and Wolf-Watz, 2006; Abbreviations: COG, Cluster of Orthologous Groups of proteins; CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats; HANK’S BALANCED SALT SOLUTION (HBSS), Hank’s Balanced Salt Solution; DMEM, Dulbecco’s Modified Eagle Medium; KEGG, Kyoto Encyclopedia of Genes and Genomes; Luria-Bertani medium supplemented with 2.5% NaCl (LB-S), Luria-Bertani medium supplemented with 2.5% (w/v) NaCl; T3SS, Type III Secretion System; V. parahaemolyticus, Vibrio parahaemolyticus; xenobiotic response element (XRE), Xenobiotic Response Element
EXPERIMENTAL DESIGN This study was comprised of two major parts: analysis of V. parahaemolyticus T3SS1 inducing conditions compared to their corresponding noninducing conditions and infection of HeLa cells in HBSS over time compared to preinfection conditions
Summary
Vibrio parahaemolyticus (V. parahaemolyticus) is an emerging food-borne bacterial pathogen that has caused significant disease in several countries. Infection is most commonly acquired by the consumption of contaminated raw or undercooked seafood, shellfish in particular, and frequently results in gastroenteritis (Yeung and Boor, 2004). This organism possesses numerous virulence factors that include two phylogenetically distinct type III secretion systems (T3SSs) located separately on each of its two chromosomes and referred to as T3SS1 and T3SS2, respectively. The T3SS is a organelle possessed by several species of Gram-negative bacteria that mediates the transport of specialized proteins, termed “effectors,” directly from the bacterial cytoplasm into the cytosol of eukaryotic cells (Galán and Wolf-Watz, 2006; Abbreviations: COG, Cluster of Orthologous Groups of proteins; CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats; HBSS, Hank’s Balanced Salt Solution; DMEM, Dulbecco’s Modified Eagle Medium; KEGG, Kyoto Encyclopedia of Genes and Genomes; LB-S, Luria-Bertani medium supplemented with 2.5% (w/v) NaCl; T3SS, Type III Secretion System; V. parahaemolyticus, Vibrio parahaemolyticus; XRE, Xenobiotic Response Element
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