Unknown,Transcriptomics,Genomics,Proteomics

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Complete genome sequence of Vibrio parahaemolyticus CHN25 and transcriptomic analysis of cold shock proteins of VpaCspA and VpaCspD in the adaptation of low temperature growth

ABSTRACT: To investigate low-temperature tolerance of the bacterium, three in-frame gene deletion mutants of VpacspA and VpacspD were constructed using homologous recombination method. When compared to the wild type strain, the growth of ΔVpacspA mutant was strongly repressed at 10 0C, whereas the deletion of VpacspD gene greatly activated the bacterium growth at the low temperature. Transcriptome data revealed that 12.4% of the expressed genes in V. parahaemolyticus CHN25 was significantly changed in ΔVpacspA mutant grown at 10 0C, including those involved in amino acid degradation, ATP-binding cassette (ABC) transporters, secretion systems, sulfur and glycerophospholipid metabolisms, whereas the low temperature elicited 10.0% of the genes from ΔVpacspD mutant, such as phosphotransferase system, nitrogen and amino acid metabolisms. Moreover, the major changed metabolic pathways in dual-gene deletion mutant (ΔVpacspAD) differed radically from those in single-gene mutants. Comparison of transcriptome profiles further revealed a number of differentially expressed genes shared among the three mutants, as well as regulators specifically, coordinately and or antagonistically regulating in the adaptation of V. parahaemolyticus CHN25 to the low-temperature growth. mRNA profiles of mid-log phase WT(G), ΔVpacspA, ΔVpacspD and ΔVpacspAD at 10 °C were generated by deep sequencing using Illumina HiSeq 2500

ORGANISM(S): Vibrio parahaemolyticus

SUBMITTER: Lanming Chen

PROVIDER: E-GEOD-65998 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The two-component system CBO0366/CBO0365 was recently demonstrated to have a role in cold tolerance of Group I Clostridium botulinum ATCC 3502. The mechanisms under its control, ultimately resulting in increased sensitivity to low temperature, are unknown. A transcriptomic analysis with DNA microarrays was performed to identify the differences in global gene expression patterns of the wild-type ATCC 3502 and a derivative mutant with insertionally inactivated cbo0365 at 37 M-BM-0C and 15 M-BM-0C. Altogether 150 or 141 chromosomal CDSs were found to be differently expressed in the cbo0365 mutant at 37 M-BM-0C or 15 M-BM-0C, respectively, and thus considered to be under direct or indirect transcriptional control of the response regulator CBO0365. Of the differentially-expressed CDSs, expression of 141 CDSs was similarly affected at both temperatures investigated, suggesting that the putative CBO0365 regulon was practically not affected by temperature. The regulon involved genes related to acetone-butanol-ethanol (ABE) fermentation, motility, to arsenic resistance, and phosphate uptake and transport. Deteriorated growth at 17 M-BM-0C was observed for mutants with disrupted ABE fermentation pathway components (crt, bdh and ctfA), arsenic detoxifying machinery components (arsC and arsR), or phosphate uptake mechanism components (phoT), suggesting roles for these mechanisms in cold tolerance of Group I C. botulinum. Electrophoretic mobility shift assays showed recombinant CBO0365 to bind to the promoter regions of crt, arsR, and phoT, as well as to the promoter region of its own operon, suggesting direct DNA-binding transcriptional activation or repression as means for CBO0365 in regulating these operons. The results provide insight to the mechanisms Group I C. botulinum utilize in coping with cold. C. botulinum ATCC 3502 cbo0365 mutant vs. wild type; 3 replicates of each strain; growth at 37C in TPGY broth batch culture and subjected to cold shock to 15C; sampling at mid-log growth phase before cold shock, and 1 h after temperature downshift to 15C (= 2 time points). Dye-swapped hybridization.