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Changes in Shewanella baltica transcriptome upon cold stress

ABSTRACT: In this work we analyzed changes in gene espression in Shewanella baltica upon cold stress. In order to imitate natural environment, we cultured bacteria in the Instant Ocean medium in a chemostat. Samples were collected after 90 and 180 minutes of cold stress and analyzed by RNA sequencing. The transcriptomic response could be characterized as global downregulation of gene expression (613 genes downregulated) and we found only 242 genes which were upregulated. We randomly tested 11 transcripts with RT-qPCR to confirm changes observed in RNA seq experiment. We observed change in expression of sigma factors namely, downregulation of the housekeeping sigma 70 encoded by rpoD and upregulation of ECF subfamily RNA polymerase sigma 24 factor rpoE2, indicating global transcription reprogramming. Analysis of metabolic pathways revealed common changes observed in other bacteria such as change in fatty acid metabolism toward synthesis of unsaturated fatty acids and upregulation of flagellar assembly genes after 90 minutes of cold stress. A novel observation was downregulation of genes encoding components of the type II secretion system. In summary,

ORGANISM(S): Shewanella baltica

PROVIDER: GSE150927 | GEO | 2020/05/21

REPOSITORIES: GEO

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Project description:Profound understanding of the mechanisms foodborne pathogenic bacteria utilize in adaptation to the environmental stress they encounter during food processing and storage is of paramount importance in design of control measures. Chill temperature is a central control measure applied in minimally processed foods; however, data on the mechanisms the foodborne pathogen Clostridium botulinum activates upon cold stress are scarce. Global gene expression analysis on the C.M-BM- botulinum ATCC 3502 strain upon temperature downshift from 37 M-BM-0C to 15M-BM- M-BM-0C was performed to identify the cold-responsive gene set of this organism. Significant up- or down-regulation of 16 and 11 genes, respectively, was observed already 1 h after the cold shock. At 5 h after the temperature downshift, 199 and 210 genes were up- and down-regulated, respectively. Thus, the cold shock rapidly affected the expression of a gene set of a relatively small size, indicating a targeted acute response to cold shock, whereas extensive metabolic remodeling took place after prolonged exposure to cold. Induction of genes related to fatty acid biosynthesis, oxidative stress response, and iron uptake and storage was observed, in addition to mechanisms previously characterized as cold-tolerance related in bacteria. Furthermore, induction of several uncharacterized DNA-binding transcriptional regulator-encoding genes was observed, suggesting involvement of novel regulatory mechanisms in the cold shock response of C.M-BM- botulinum. The role of such regulatory proteins, CBO0477 and CBO0558A, in cold tolerance of C.M-BM- botulinum ATCC 3502 was demonstrated by the deteriorated growth of mutants of the respective genes at 17M-BM- M-BM-0C. C. botulinum ATCC 3502 wild type cold-shocked vs. pre-cold-shock; 3 replicates; 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 and 5 h after temperature downshift to 15C (= 3 time points). Dye-swapped hybridization.

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Changes in Shewanella baltica transcriptome upon cold stress

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