Gene expression of Vibrio parahaemolyticus growing in laboratory isolation conditions compared to those common in its natural ocean environment

ABSTRACT In this work, we describe a new gene controlling lateral flagellar gene expression. The gene encodes ScrG, a protein containing GGDEF and EAL domains. This is the second GGDEF-EAL-encoding locus determined to be involved in the regulation of swarming: the first was previously characterized and named scrABC (for “swarming and capsular polysaccharide regulation”). GGDEF and EAL domain-containing proteins participate in the synthesis and degradation of the nucleotide signal cyclic di-GMP (c-di-GMP) in many bacteria. Overexpression of scrG was sufficient to induce lateral flagellar gene expression in liquid, decrease biofilm formation, decrease cps gene expression, and suppress the ΔscrABC phenotype. Removal of its EAL domain reversed ScrG activity, converting ScrG to an inhibitor of swarming and activator of cps expression. Overexpression of scrG decreased the intensity of a 32P-labeled nucleotide spot comigrating with c-di-GMP standard, whereas overexpression of scrG Δ EAL enhanced the intensity of the spot. Mutants with defects in scrG showed altered swarming and lateral flagellin production and colony morphology (but not swimming motility); furthermore, mutation of two GGDEF-EAL-encoding loci (scrG and scrABC) produced cumulative effects on swarming, lateral flagellar gene expression, lateral flagellin production and colony morphology. Mutant analysis supports the assignment of the primary in vivo activity of ScrG to acting as a phosphodiesterase. The data are consistent with a model in which multiple GGDEF-EAL proteins can influence the cellular nucleotide pool: a low concentration of c-di-GMP favors surface mobility, whereas high levels of this nucleotide promote a more adhesive Vibrio parahaemolyticus cell type.

Download Full-text

PVv3, a New Shuttle Vector for Gene Expression in Vibrio vulnificus

Applied and Environmental Microbiology ◽

10.1128/aem.03720-13 ◽

2013 ◽

Vol 80 (4) ◽

pp. 1477-1481 ◽

Cited By ~ 8

Author(s):

Karina Klevanskaa ◽

Nadja Bier ◽

Kerstin Stingl ◽

Eckhard Strauch ◽

Stefan Hertwig

Keyword(s):

Gene Expression ◽

Escherichia Coli ◽

Green Fluorescent Protein ◽

Vibrio Parahaemolyticus ◽

Fluorescent Protein ◽

Vibrio Vulnificus ◽

Shuttle Vector ◽

Content Type ◽

E Coli ◽

Green Fluorescent

ABSTRACTAn efficient electroporation procedure forVibrio vulnificuswas designed using the new cloning vector pVv3 (3,107 bp). Transformation efficiencies up to 2 × 106transformants per μg DNA were achieved. The vector stably replicated in bothV. vulnificusandEscherichia coliand was also successfully introduced intoVibrio parahaemolyticusandVibrio cholerae. To demonstrate the suitability of the vector for molecular cloning, the green fluorescent protein (GFP) gene and thevvhBAhemolysin operon were inserted into the vector and functionally expressed inVibrioandE. coli.

Download Full-text

The mechanisms that regulate Vibrio parahaemolyticus virulence gene expression differ between pathotypes

Microbial Genomics ◽

10.1099/mgen.0.000182 ◽

2018 ◽

Vol 4 (6) ◽

Cited By ~ 2

Author(s):

Nicholas Petronella ◽

Jennifer Ronholm

Keyword(s):

Gene Expression ◽

Vibrio Parahaemolyticus ◽

Virulence Gene ◽

Virulence Gene Expression

Download Full-text

Metabolome response to temperature-induced virulence gene expression in two genotypes of pathogenic Vibrio parahaemolyticus

BMC Microbiology ◽

10.1186/s12866-016-0688-5 ◽

2016 ◽

Vol 16 (1) ◽

Cited By ~ 6

Author(s):

Bo Feng ◽

Zhuoran Guo ◽

Weijia Zhang ◽

Yingjie Pan ◽

Yong Zhao

Keyword(s):

Gene Expression ◽

Vibrio Parahaemolyticus ◽

Virulence Gene ◽

Virulence Gene Expression ◽

Pathogenic Vibrio ◽

Response To Temperature

Download Full-text

Differential gene expression and extracellular secretion of the collagenolytic enzymes by the pathogen Vibrio parahaemolyticus

FEMS Microbiology Letters ◽

10.1111/j.1574-6968.2008.01159.x ◽

2008 ◽

Vol 283 (2) ◽

pp. 176-181 ◽

Cited By ~ 18

Author(s):

Shin-ichi Miyoshi ◽

Yuko Nitanda ◽

Kaori Fujii ◽

Kiyomi Kawahara ◽

Tao Li ◽

...

Keyword(s):

Gene Expression ◽

Differential Gene Expression ◽

Vibrio Parahaemolyticus ◽

Extracellular Secretion ◽

Differential Gene

Download Full-text

Dietary effect of Sargassum wightii fucoidan to enhance growth, prophenoloxidase gene expression of Penaeus monodon and immune resistance to Vibrio parahaemolyticus

Fish & Shellfish Immunology ◽

10.1016/j.fsi.2014.05.037 ◽

2014 ◽

Vol 39 (2) ◽

pp. 439-449 ◽

Cited By ~ 26

Author(s):

Madasamy Sivagnanavelmurugan ◽

Bergmans Jude Thaddaeus ◽

Arunachalam Palavesam ◽

Grasian Immanuel

Keyword(s):

Gene Expression ◽

Vibrio Parahaemolyticus ◽

Penaeus Monodon ◽

Sargassum Wightii ◽

Immune Resistance

Download Full-text

DNA-Binding Protein HU Coordinates Pathogenicity in Vibrio parahaemolyticus

Journal of Bacteriology ◽

10.1128/jb.00306-15 ◽

2015 ◽

Vol 197 (18) ◽

pp. 2958-2964 ◽

Cited By ~ 9

Author(s):

Ngoc Quang Phan ◽

Takashi Uebanso ◽

Takaaki Shimohata ◽

Mutsumi Nakahashi ◽

Kazuaki Mawatari ◽

...

Keyword(s):

Gene Expression ◽

Growth Rate ◽

Virulence Factors ◽

Vibrio Parahaemolyticus ◽

Binding Protein ◽

Negative Regulator ◽

Independent Manner ◽

Content Type ◽

Expression Levels ◽

Hu Proteins

ABSTRACTHU is one of the most abundant nucleoid-associated proteins in bacterial cells and regulates the expression of many genes involved in growth, motility, metabolism, and virulence. It is known thatVibrio parahaemolyticuspathogenicity is related to its characteristic rapid growth and that type III secretion system 1 (T3SS1) contributes to its cytotoxicity. However, it is not known if HU plays a role in the pathogenicity ofV. parahaemolyticus. In the present study, we investigated the effect of HU proteins HU-2 (HUα) (V. parahaemolyticus2911 [vp2911]) and HUβ (vp0920) on the pathogenicity ofV. parahaemolyticus. We found that a deletion of both HU subunits (yielding the ΔHUs [Δvp0920Δvp2911] strain), but not single deletions, led to a reduction of the growth rate. In addition, expression levels of T3SS1-related genes, includingexsA(positive regulator),exsD(negative regulator),vp1680(cytotoxic effector), andvp1671(T3SS1 apparatus), were reduced in the ΔHUs strain compared to the wild type (WT). As a result, cytotoxicity to HeLa cells was decreased in the ΔHUs strain. The additional deletion ofexsDin the ΔHUs strain restored T3SS1-related gene expression levels and cytotoxicity but not the growth rate. These results suggest that the HU protein regulates the levels of T3SS1 gene expression and cytotoxicity in a growth rate-independent manner.IMPORTANCENucleoid-binding protein HU regulates cellular behaviors, including nucleoid structuring, general recombination, transposition, growth, replication, motility, metabolism, and virulence. It is thought that both the number of bacteria and the number of virulence factors may affect the pathogenicity of bacteria. In the present study, we investigated which factor(s) has a dominant role during infection in one of the most rapidly growing bacterial species,Vibrio parahaemolyticus. We found thatV. parahaemolyticuscytotoxicity is regulated, in a growth rate-independent manner, by the HU proteins through regulation of a number of virulence factors, including T3SS1 gene expression.

Download Full-text