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Analysis of tRNA ^Cys processing under salt stress in Bacillus subtilis spore outgrowth using RNA sequencing data.

ABSTRACT: Background: In spore-forming bacteria, the molecular mechanisms of accumulation of transfer RNA (tRNA) during sporulation must be a priority as tRNAs play an essential role in protein synthesis during spore germination and outgrowth. However, tRNA processing has not been extensively studied in these conditions, and knowledge of these mechanisms is important to understand long-term stress survival. Methods:To gain further insight into tRNA processing during spore germination and outgrowth, the expression of the single copy tRNA ^Cys gene was analyzed in the presence and absence of 1.2 M NaCl in Bacillus subtilis using RNA-Seq data obtained from the Gene Expression Omnibus (GEO) database. The CLC Genomics work bench 12.0.2 (CLC Bio, Aarhus, Denmark, https://www.qiagenbioinformatics.com/) was used to analyze reads from the tRNA ^Cys gene. Results:The results show that spores store different populations of tRNA ^Cys-related molecules. One such population, representing 60% of total tRNA ^Cys, was composed of tRNA ^Cys fragments. Half of these fragments (3´-tRF) possessed CC, CCA or incorrect additions at the 3´end. tRNA ^Cys with correct CCA addition at the 3´end represented 23% of total tRNA ^Cys, while with CC addition represented 9% of the total and with incorrect addition represented 7%. While an accumulation of tRNA ^Cys precursors was induced by upregulation of the rrnD operon under the control of σ ^A -dependent promoters under both conditions investigated, salt stress produced only a modest effect on tRNA ^Cys expression and the accumulation of tRNA ^Cys related species. Conclusions:The results demonstrate that tRNA ^Cys molecules resident in spores undergo dynamic processing to produce functional molecules that may play an essential role during protein synthesis.

SUBMITTER: Arvizu Hernandez I

PROVIDER: S-EPMC8097732 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Changes with time of a population of Bacillus weihenstephanensis KBAB4 and Bacillus licheniformis AD978 dormant spores into germinated spores and vegetative cells were followed by flow cytometry, at pH ranges of 4.7 to 7.4 and temperatures of 10°C to 37°C for B. weihenstephanensis and 18°C to 59°C for B. licheniformis Incubation conditions lower than optimal temperatures or pH led to lower proportions of dormant spores able to germinate and extended time of germination, a lower proportion of germinated spores able to outgrow, an extension of their times of outgrowth, and an increase of the heterogeneity of spore outgrowth time. A model based on the strain growth limits was proposed to quantify the impact of incubation temperature and pH on the passage through each physiological stage. The heat treatment temperature or time acted independently on spore recovery. Indeed, a treatment at 85°C for 12?min or at 95°C for 2?min did not have the same impact on spore germination and outgrowth kinetics of B. weihenstephanensis despite the fact that they both led to a 10-fold reduction of the population. Moreover, acidic sporulation pH increased the time of outgrowth 1.2-fold and lowered the proportion of spores able to germinate and outgrow 1.4-fold. Interestingly, we showed by proteomic analysis that some proteins involved in germination and outgrowth were detected at a lower abundance in spores produced at pH 5.5 than in those produced at pH 7.0, maybe at the origin of germination and outgrowth behavior of spores produced at suboptimal pH.IMPORTANCE Sporulation and incubation conditions have an impact on the numbers of spores able to recover after exposure to sublethal heat treatment. Using flow cytometry, we were able to follow at a single-cell level the changes in the physiological states of heat-stressed spores of Bacillus spp. and to discriminate between dormant spores, germinated spores, and outgrowing vegetative cells. We developed original mathematical models that describe (i) the changes with time of the proportion of cells in their different states during germination and outgrowth and (ii) the influence of temperature and pH on the kinetics of spore recovery using the growth limits of the tested strains as model parameters. We think that these models better predict spore recovery after a sublethal heat treatment, a common situation in food processing and a concern for food preservation and safety.

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Analysis of tRNA ^Cys processing under salt stress in Bacillus subtilis spore outgrowth using RNA sequencing data.

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Dataset Information

Analysis of tRNA Cys processing under salt stress in Bacillus subtilis spore outgrowth using RNA sequencing data.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Analysis of tRNA ^Cys processing under salt stress in Bacillus subtilis spore outgrowth using RNA sequencing data.