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SpoVG is Necessary for Sporulation in Bacillus anthracis.

ABSTRACT: 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 ?spoVG 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 ?-galactosidase activity assays. The effects on sporulation of the ?spoVG 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 ?spoVG strain with spoIIBBs, the resulting strain pBspoIIBBs/?spoVG 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/?spoVG compared with the ?spoVG strain. Transcription of the coat development gene cotE was similar in the pBspoIIBBs/?spoVG and ?spoVG 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.

SUBMITTER: Chen M

PROVIDER: S-EPMC7232415 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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SpoVG is Necessary for Sporulation in Bacillus anthracis.

Chen Meng M Lyu Yufei Y Feng Erling E Zhu Li L Pan Chao C Wang Dongshu D Liu Xiankai X Wang Hengliang H

Microorganisms 20200410 4

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 demonst ...[more]

PMID: 32290166

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Project description:BackgroundBacillus cereus 0-9, a Gram-positive, endospore-forming bacterium isolated from healthy wheat roots in our previous research, is considered to be an effective biocontrol strain against several soil-borne plant diseases. SpoVG, a regulator that is broadly conserved among many Gram-positive bacteria, may help this organism coordinate environmental growth and virulence to survive. This study aimed to explore the multiple functions of SpoVG in B. cereus 0-9.MethodsThe gene knockout strains were constructed by homologous recombination, and the sporulation process of B. cereus 0-9 and its mutants were observed by fluorescence staining method. We further determined the spore yields and biofilm formation abilities of test strains. Transcriptional fusion strains were constructed by overlapping PCR technique, and the promoter activity of the target gene was detected by measuring its fluorescence intensity. The biofilm production and colonial morphology of B. cereus 0-9 and its mutants were determined to study the functions of the target genes, and the transcription level of the target gene was determined by qRT-PCR.ResultsAccording to observation of the sporulation process of B. cereus 0-9 in germination medium, SpoVG is crucial for regulating sporulation stage V of B. cereus 0-9, which is identical to that of Bacillus subtilis but differs from that of Bacillus anthracis. In addition, SpoVG could influence biofilm formation of B. cereus 0-9. The transcription levels of two genes closely related to biofilm-formation, sipW and calY, were downregulated in a ΔspoVG mutant. The role of SpoVG in regulating biofilm formation was further explored by deleting the genes abrB and sinR in the ΔspoVG mutant, respectively, generating the double mutant strains ΔspoVGΔabrB and ΔspoVGΔsinR. The phenotypes of these double mutants were congruent with those of the single abrB and sinR deletion strains, respectively, which showed increased biofilm formation. This indicated that spoVG was located upstream of abrB and sinR in the regulatory pathway of B. cereus biofilm formation. Further, the results of qRT-PCR and the luminescence intensity of transcriptional fusion strains indicated that spoVG gene deletion could inhibit the transcription of Spo0A.ConclusionsSpoVG, an important regulator in the sporulation of B. cereus, is located upstream of Spo0A and participates in regulation of biofilm formation of B. cereus 0-9 through regulating the transcription level of spo0A. Sporulation and biofilm formation are crucial mechanisms by which bacteria respond to adverse conditions. SpoVG is therefore an important regulator of Spo0A and is crucial for both sporulation and biofilm formation of B. cereus 0-9. This study provides a new insight into the regulatory mechanism of environmental adaptation in bacteria and a foundation for future studies on biofilm formation of B. cereus.

Dataset Information

SpoVG is Necessary for Sporulation in Bacillus anthracis.

Publications

SpoVG is Necessary for Sporulation in <i>Bacillus anthracis</i>.

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