Independence of Bacillus subtilis spore outgrowth from DNA synthesis

Ginsberg, Dalia; Keynan, A.

doi:10.1128/jb.136.1.111-116.1978

Cited by 13 publications

(4 citation statements)

References 19 publications

(24 reference statements)

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“…It is well known that water from the environment enters the dehydrated spore core during the initial phases of spore germination, thereby reactivating the latent processes within the spore. Characteristically, that mRNA synthesis and protein synthesis are the primary processes to be activated was demonstrated long ago by Ginsberg and Keynan ( 30 ). In agreement with this observation, our data confirm that the transcription machinery is highly active beyond germination, with the expression of ∼2,400 mRNA transcripts being upregulated.…”

Section: Discussionmentioning

confidence: 97%

Integrative Analysis of Proteome and Transcriptome Dynamics during Bacillus subtilis Spore Revival

Swarge

Abhyankar

Jonker

et al. 2020

mSphere

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Bacillus subtilis spores can reactivate their metabolism through germination upon contact with germinants and can develop into vegetative cells upon outgrowth. However, the mechanisms at the basis of the molecular machinery that triggers the spore germination and outgrowth processes are still largely unclear. To gain further insights into these processes, the transcriptome and proteome changes occurring during the conversion of spores to vegetative cells were analyzed in the present study. For each time point sampled, the changes in the spore proteome were quantitatively monitored relative to the proteome of metabolically 15N-labeled vegetative cells. Of the quantified proteins, 60% are shared by vegetative cells and spores, indicating that the spores have a minimal protein set, sufficient to resume metabolism upon completion of germination. These shared proteins thus represent the most basic “survival kit” for spore-based life. We observed no significant change in the proteome or the transcriptome until the spore’s completion of germination. Our analysis identified 34 abundant mRNA transcripts in the dormant spores, 31 of which are rapidly degraded after germination. In outgrowing spores, we identified 3,152 differentially expressed genes and have demonstrated the differential expression of 322 proteins with our mass spectrometry analyses. Our data also showed that 173 proteins from dormant spores, including both proteins unique to spores and proteins shared with vegetative cells, were lost after completion of germination. The observed diverse timings of synthesis of different protein sets in spore outgrowth revealed a putative core strategy underlying the revival of ‘life’ from the B. subtilis spore. IMPORTANCE This study demonstrated the progress of macromolecular synthesis during Bacillus subtilis spore germination and outgrowth. The transcriptome analysis has additionally allowed us to trace gene expression during this transformation process. For the first time, the basic survival kit for spore-based life has been identified. In addition, in this analysis based on monitoring of protein levels in germinating and outgrowing spores, the transition from (ribo)nucleotide and amino acid biosynthesis to the restoration of all metabolic pathways can be clearly seen. The integrative multi-omics approach applied in this study thus has helped us to achieve a comprehensive overview of the molecular mechanisms at the basis of spore germination and outgrowth as well as to identify important knowledge gaps in need of further study.

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Section: Discussionmentioning

confidence: 97%

Integrative Analysis of Proteome and Transcriptome Dynamics during Bacillus subtilis Spore Revival

Swarge

Abhyankar

Jonker

et al. 2020

mSphere

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“…Nalidixic acid and novobiocin are known to act differentially on germinating spores, on vegeta-tive cells, and on the initiation of sporulation (8,9); hence, studies of DNA gyrase as it influences the state of the chromosome are justified in attempts to explain the events important in regulating the differentiated state.…”

mentioning

confidence: 99%

Bacillus subtilis deoxyribonucleic acid gyrase

Sugino¹,

Bott²

1980

J Bacteriol

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Bacillus subtilis 168 was shown to contain a deoxyribonucleic acid (DNA) gyrase activity which closely resembled those of the enzymes isolated from Escherichia coli and Micrococcus luteus in its enzymatic requirements, substrate specificity, and sensitivity to several antibiotics. The enzyme was purified from the wild type and nalidixic acid-resistant and novobiocin-resistant mutants of B. subtilis and was functionally characterized in vitro. The genetic loci nalA and novA but not novB were shown to code for portions of the functional gyrase. Enzyme from the antibiotic-resistant mutants was resistant to the drug in vitro. The most striking observation was the remarkable similarity between the B. subtilis enzyme and other DNA gyrases, especially with respect to the oxolinic acid-induced DNA cleavage in the presence of sodium dodecyl sulfate. All of the enzymes appeared to possess the same specificity of cutting sites regardless of the source or type of DNA used. This result implies that gyrase binding to DNA is highly specific.

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“…It is well known that during the initial phases of spore germination, the water from the environment enters the dehydrated spore core thereby reactivating the latent processes within the spore. Characteristically, the mRNA and protein synthesis are the primary processes to be activated as was already demonstrated long time ago by Ginsberg and Keynan 30 . In agreement with this observation, our data confirms that the transcription machinery is highly active during and beyond the phase transition period with the expression of ∼ 2400 mRNA transcripts being upregulated.…”

Section: Discussionmentioning

confidence: 84%

Integrative analysis of proteome and transcriptome dynamics duringBacillus subtilisspore revival

Swarge

Jonker

Abhyankar

et al. 2019

Preprint

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Bacillus subtilis forms highly resistant, metabolically inactive dormant spores upon nutrient limitation. These endospores pose challenges to the food and medical sectors. Spores reactivate their metabolism upon contact with germinants and develop into vegetative cells.The activation of the molecular machinery that triggers the progress of germination and spore outgrowth is still unsettled. To gain further insight in spore germination and outgrowth processes, the transcriptome and proteome changeover during spore germination and outgrowth to vegetative cells, was analysed. B. subtilis transcriptome analysis allow us to trace the different functional groups of genes expressed. For each time-point sample, the change in the spore proteome was quantitatively monitored relative to the reference proteome of 15 N metabolically labelled vegetative cells. We observed until the phase transition, i.e. completion of germination, no significant change in the proteome. We have identified 36 transcripts present abundantly in the dormant spores. This number is in close agreement with the previous findings. These transcripts mainly belong to the genes encoding small acid soluble proteins (sspE, sspO, sspI, sspK, sspF) and proteins with uncharacterized functions.We observed in total 3152 differentially expressed genes, but 'only' 323 differentially expressed proteins (total 451 proteins identified and quantified). Our data shows that 173 proteins from dormant spores, both spore unique proteins and protein shared with vegetative cells, are lost during the phase transitioning period. This loss is in addition to the active protein degradation, undertaken by the spore proteases such as Gpr, as germination and outgrowth proceeds. Further analysis is required to functionally interpret the observed protein loss. The observed diverse timing of the synthesis of different protein sets reveals a putative core-strategy of the revival of 'life' starting from the B. subtilis spore.

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Independence of Bacillus subtilis spore outgrowth from DNA synthesis

Cited by 13 publications

References 19 publications

Integrative Analysis of Proteome and Transcriptome Dynamics during Bacillus subtilis Spore Revival

Integrative Analysis of Proteome and Transcriptome Dynamics during Bacillus subtilis Spore Revival

Bacillus subtilis deoxyribonucleic acid gyrase

Integrative analysis of proteome and transcriptome dynamics duringBacillus subtilisspore revival

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