Identification of a new developmental locus in Bacillus subtilis by construction of a deletion mutation in a cloned gene under sporulation control

Rosenbluh, A; Banner, Carl; Losick, Richard; Fitz-James, Philip C.

doi:10.1128/jb.148.1.341-351.1981

Cited by 61 publications

(25 citation statements)

References 22 publications

(22 reference statements)

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“…Although the absence of spoVG causes little impairment in sporulation in the model organism B. subtilis, it caused aberrations in the cortex [7,16] and amplified the effects of a spoIIB mutation [5,15]. To investigate whether SpoVG influenced sporulation in B. anthracis A16R, a spoVG mutant (∆spoVG) and a plasmid-based complementation strain (R∆spoVG) were constructed.…”

Section: Deletion Of Spovg Results In a Spore Formation Defect In B mentioning

confidence: 99%

SpoVG Is Necessary for Sporulation in Bacillus anthracis

Chen¹,

Lyu²,

Feng³

et al. 2020

Microorganisms

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The Bacillus anthracis spore constitutes the infectious form of the bacterium, and sporulation is an important process in the organism’s life cycle. Herein, we show that disruption of SpoVG resulted in defective B. anthracis sporulation. Confocal microscopy demonstrated that a ΔspoVG mutant could not form an asymmetric septum, the first morphological change observed during sporulation. Moreover, levels of spoIIE mRNA were reduced in the spoVG mutant, as demonstrated using β-galactosidase activity assays. The effects on sporulation of the ΔspoVG mutation differed in B. anthracis from those in B. subtilis because of the redundant functions of SpoVG and SpoIIB in B. subtilis. SpoVG is highly conserved between B. anthracis and B. subtilis. Conversely, BA4688 (the protein tentatively assigned as SpoIIB in B. anthracis) and B. subtilis SpoIIB (SpoIIBBs) share only 27.9% sequence identity. On complementation of the B. anthracis ΔspoVG strain with spoIIBBs, the resulting strain pBspoIIBBs/ΔspoVG could not form resistant spores, but partially completed the prespore engulfment stage. In agreement with this finding, mRNA levels of the prespore engulfment gene spoIIM were significantly increased in strain pBspoIIBBs/ΔspoVG compared with the ΔspoVG strain. Transcription of the coat development gene cotE was similar in the pBspoIIBBs/ΔspoVG and ΔspoVG strains. Thus, unlike in B. subtilis, SpoVG appears to be required for sporulation in B. anthracis, which provides further insight into the sporulation mechanisms of this pathogen.

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Section: Deletion Of Spovg Results In a Spore Formation Defect In B mentioning

confidence: 99%

SpoVG Is Necessary for Sporulation in Bacillus anthracis

Chen¹,

Lyu²,

Feng³

et al. 2020

Microorganisms

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“…Unfortunately, fragment 14 in pPL714 did not specify any additional detectable host phenotype. One approach to assigning a genetic function to fragment 14 involves introducing a deletion within the fragment into the B. subtilis chromosome and observing the phenotypic variation in the host cell (22). This has been tested by recombining a derivative of fragment 14, deleted for an internal 2-kb BclI-HpaI segment (see Fig.…”

Section: Discussionmentioning

confidence: 99%

Restriction fragments that exert promoter activity during postexponential growth of Bacillus subtilis

Mongkolsuk¹,

Chiang²,

Reynolds³

et al. 1983

J Bacteriol

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Two restriction fragments of Bacillus subtilis DNA were identified which caused the cat-86 gene present on the promoter cloning plasmid pPL703 to be activated predominantly during postexponential growth of host cells. The postexponential increase was observed in both sporulation-positive strains and in a spoOA mutant of B. subtilis. However, the postexponential increase in the cat-86 gene product, chloramphenicol acetyltransferase, was diminished or not observed when the plasmid-containing cells were grown in the presence of excess glucose. The promoter-containing fragment, designated as 33, was mapped to a site on the B. subtilis chromosome adjacent to hisA. The other fragment, 14, mapped to a site adjacent to ctrA. When present on a high-copy vector, both fragments caused a reduction in the sporulation frequency of host cells. Fragment 33 in high copy number conferred on B. subtilis cells three additional phenotypic changes: brown colony color, intracellular inclusions, and, in a protease-deficient mutant, the production of extracellular protease activity. These activities were observed only in postexponential-phase cultures.

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“…Since H is active in a citC mutant (28), the mechanism by which the absence of ICDH activity interferes with asymmetric septation is not known. As shown in the accompanying paper (36), further analysis of the stage I block in a citC mutant has revealed that inactivation of the spoVG gene (42,44,46) allows the mutant cells to overcome in part their defect in asymmetric septation and activate forespore-specific, F -dependent genes. SpoVG, which is induced at the onset of stationary phase (54), acts as a transient negative regulator of the pathway leading to asymmetric septum formation (36).…”

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confidence: 98%

Metabolic Imbalance and Sporulation in an Isocitrate Dehydrogenase Mutant of Bacillus subtilis

et al. 1999

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A Bacillus subtilis mutant with a deletion in thecitC gene, encoding isocitrate dehydrogenase, the third enzyme of the tricarboxylic acid branch of the Krebs cycle, exhibited reduced growth yield in broth medium and had greatly reduced ability to sporulate compared to the wild type due to a block at stage I, i.e., a failure to form the polar division septum. In early stationary phase, mutant cells accumulated intracellular and extracellular concentrations of citrate and isocitrate that were at least 15-fold higher than in wild-type cells. The growth and sporulation defects of the mutant could be partially bypassed by deletion of the major citrate synthase gene (citZ), by raising the pH of the medium, or by supplementation of the medium with certain divalent cations, suggesting that abnormal accumulation of citrate affects survival of stationary-phase cells and sporulation by lowering extracellular pH and chelating metal ions. While these genetic and environmental alterations were not sufficient to allow the majority of the mutant cell population to pass the stage I block (lack of asymmetric septum formation), introduction of the sof-1 mutant form of the Spo0A transcription factor, when coupled with a reduction in citrate synthesis, restored sporulation gene expression and spore formation nearly to wild-type levels. Thus, the primary factor inhibiting sporulation in a citC mutant is abnormally high accumulation of citrate, but relief of this metabolic defect is not by itself sufficient to restore competence for sporulation.

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Identification of a new developmental locus in Bacillus subtilis by construction of a deletion mutation in a cloned gene under sporulation control

Cited by 61 publications

References 22 publications

SpoVG Is Necessary for Sporulation in Bacillus anthracis

SpoVG Is Necessary for Sporulation in Bacillus anthracis

Restriction fragments that exert promoter activity during postexponential growth of Bacillus subtilis

Metabolic Imbalance and Sporulation in an Isocitrate Dehydrogenase Mutant of Bacillus subtilis

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