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Evidence that Bacillus subtilis sporulation induced by the stringent response is caused by the decrease in GTP or

K Ochi, J Kandala, E Freese

    Journal of Bacteriology
    |August 1, 1982
    PubMed
    Summary
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    Bacillus subtilis sporulation is triggered by a decrease in GTP levels, not an increase in ppGpp and pppGpp. This finding challenges the traditional view of the stringent response in bacterial development.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Bacterial Physiology

    Background:

    • The stringent response in Bacillus subtilis is a key survival mechanism during nutrient limitation.
    • This response is traditionally associated with the accumulation of alarmones like guanosine-5'-diphosphate-3'-diphosphate (ppGpp) and guanosine-5'-triphosphate-3'-diphosphate (pppGpp).
    • The role of these nucleotides in initiating sporulation, a complex differentiation process, has been extensively studied.

    Purpose of the Study:

    • To investigate the precise molecular mechanism by which partial amino acid deprivation triggers sporulation in Bacillus subtilis.
    • To determine whether the increase in ppGpp and pppGpp or the decrease in GTP is the primary driver of sporulation initiation.
    • To elucidate the role of GTP levels in the stringent response and subsequent sporulation.

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    Main Methods:

    • Utilized a mutant Bacillus subtilis strain (Myc) with partial resistance to mycophenolate, an inhibitor of IMP dehydrogenase.
    • Analyzed intracellular nucleotide levels (GTP, ppGpp, pppGpp) under conditions of partial amino acid deprivation.
    • Assessed sporulation efficiency in the wild-type and mutant strains with and without the addition of mycophenolate or decoyinine.

    Main Results:

    • The Myc mutant exhibited an increase in ppGpp and pppGpp upon amino acid deprivation but a significantly reduced decrease in GTP.
    • Sporulation was minimal in the Myc mutant, indicating that the increase in ppGpp/pppGpp alone is insufficient for robust sporulation.
    • Addition of mycophenolate or decoyinine, which further decreased GTP levels, restored extensive sporulation in the Myc mutant.

    Conclusions:

    • The initiation of Bacillus subtilis sporulation under partial amino acid deprivation is primarily driven by a decrease in GTP levels, not the accumulation of ppGpp and pppGpp.
    • This finding refines the understanding of the stringent response pathway and its role in bacterial differentiation.
    • GTP availability is a critical regulatory factor in the transition from vegetative growth to sporulation.