Surfaces of Spo0A and RNA Polymerase Sigma Factor A That Interact at the
            <i>spoIIG</i>
            Promoter in
            <i>Bacillus subtilis</i>

Kumar, Amrita; Starke, Cindy Buckner; DeZalia, Mark; Moran, Charles P.

doi:10.1128/jb.186.1.200-206.2004

Cited by 13 publications

(16 citation statements)

References 31 publications

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“…The barrier posed by the long spacer could be overcome in at least two ways. First, a transcription factor could bind to and bend spacer DNA (2) or bind to the promoter Ϫ35 region and interact with region 4 of A in such a way that its interaction with the Ϫ10 region is improved (52). If such a transcription factor was developmentally regulated, this could explain developmental regulation of dev transcription.…”

Section: Discussionmentioning

confidence: 99%

Regulation of dev , an Operon That Includes Genes Essential for Myxococcus xanthus Development and CRISPR-Associated Genes and Repeats

et al. 2007

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Expression of dev genes is important for triggering spore differentiation inside Myxococcus xanthus fruiting bodies. DNA sequence analysis suggested that dev and cas (CRISPR-associated) genes are cotranscribed at the dev locus, which is adjacent to CRISPR (clustered regularly interspaced short palindromic repeats). Analysis of RNA from developing M. xanthus confirmed that dev and cas genes are cotranscribed with a short upstream gene and at least two repeats of the downstream CRISPR, forming the dev operon. The operon is subject to strong, negative autoregulation during development by DevS. The dev promoter was identified. Its ؊35 and ؊10 regions resemble those recognized by M. xanthus A RNA polymerase, the homolog of Escherichia coli 70 , but the spacer may be too long (20 bp); there is very little expression during growth. Induction during development relies on at least two positive regulatory elements located in the coding region of the next gene upstream. At least two positive regulatory elements and one negative element lie downstream of the dev promoter, such that the region controlling dev expression spans more than 1 kb. The results of testing different fragments for dev promoter activity in wild-type and devS mutant backgrounds strongly suggest that upstream and downstream regulatory elements interact functionally. Strikingly, the 37-bp sequence between the two CRISPR repeats that, minimally, are cotranscribed with dev and cas genes exactly matches a sequence in the bacteriophage Mx8 intP gene, which encodes a form of the integrase needed for lysogenization of M. xanthus.

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Section: Discussionmentioning

confidence: 99%

Regulation of dev , an Operon That Includes Genes Essential for Myxococcus xanthus Development and CRISPR-Associated Genes and Repeats

et al. 2007

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“…The best-understood mechanism of promoter activation by Spo0A is the activation of the high-threshold, A -dependent spoIIG promoter (4,12,(19)(20)(21)23). In this promoter, sequences similar to the Ϫ10 and Ϫ35 hexamers, which signal recognition of promoters by A -RNA polymerase, are separated by 22 bp, a distance that is ordinarily considered too great for productive interaction of these sequences with A (15).…”

Section: Discussionmentioning

confidence: 99%

“…At this promoter, the consensus Ϫ35-like promoter element is centered between positions Ϫ37 and Ϫ36 and is overlapped by a Spo0A binding site (20). Based on molecular modeling and genetic assays, it has been proposed that binding of Spo0AϳP occludes binding of A region 4 to the consensus Ϫ35-like sequence and interaction between Spo0A and A positions in region 4 of A 18 bp upstream from the Ϫ10 region of the spoIIG promoter (12). This positioning of A allows region 2 of A to interact productively with the Ϫ10 region of the spoIIG promoter, thus stimulating its activity.…”

Section: Discussionmentioning

confidence: 99%

“…Integration of pBG1PLK-SKF into the wild-type B. subtilis chromosomal amyE locus by homologous recombination was confirmed by testing for loss of amylase production by iodine staining and by PCR and DNA sequencing. The resulting strain was later transformed with ⌬abrB and/or ⌬spo0A knockout plasmids (12), producing corresponding B. subtilis strains (Table 1). Overlapping PCR was used to introduce base pair substitutions in the wildtype skf promoter.…”

Section: Methodsmentioning

confidence: 99%

“…The sequence of the skf promoter, which is activated by low levels of Spo0AϳP (9), appears to indicate that Spo0AϳP may directly activate this promoter by a mechanism that is different from that of the spoIIG promoter, which has been studied more extensively (4,12,(19)(20)(21)23). Comparison of the relative affinities of Spo0A for the high-and low-threshold promoters shows that spoIIG has a K d of 1,700 nM whereas the skf promoter has an apparent K d value of 26 nM (9).…”

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confidence: 99%

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Spo0A-Dependent Activation of an Extended −10 Region Promoter in Bacillus subtilis

et al. 2006

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At the onset of endospore formation in Bacillus subtilis the DNA-binding protein Spo0A directly activates transcription from promoters of about 40 genes. One of these promoters, Pskf, controls expression of an operon encoding a killing factor that acts on sibling cells. AbrB-mediated repression of Pskf provides one level of security ensuring that this promoter is not activated prematurely. However, Spo0A also appears to activate the promoter directly, since Spo0A is required for Pskf activity in a ⌬abrB strain. Here we investigate the mechanism of Pskf activation. DNase I footprinting was used to determine the locations at which Spo0A bound to the promoter, and mutations in these sites were found to significantly reduce promoter activity. The sequence near the ؊10 region of the promoter was found to be similar to those of extended ؊10 region promoters, which contain a TRTGn motif. Mutational analysis showed that this extended ؊10 region, as well as other base pairs in the ؊10 region, is required for Spo0A-dependent activation of the promoter. We found that a substitution of the consensus base pair for the nonconsensus base pair at position ؊9 of Pskf produced a promoter that was active constitutively in both ⌬abrB and ⌬spo0A ⌬abrB strains. Therefore, the base pair at position ؊9 of Pskf makes its activity dependent on Spo0A binding, and the extended ؊10 region motif of the promoter contributes to its high level of activity.

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Molecular insights into the initiation of sporulation in Gram-positive bacteria: new technologies for an old phenomenon

Stephenson

Lewis

2005

FEMS Microbiology Reviews

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The last decade has witnessed extensive, and widespread, changes in scientific technologies that have impacted significantly upon the study of the life sciences. Arguably, the biggest advances in our comprehension of simple and complex biological processes have come as a consequence of obtaining the complete DNA sequence of organisms. It is likely that we will become accustomed to hearing of quantum leaps in the study and understanding of the biology of higher eukaryotes in the coming years, now that (near) complete genome sequences are available for man, mouse and rat. In this review, we will discuss the impact of genome sequence data, and the use of new scientific technologies that have emerged largely as consequence of the availability of this information, on the study of the master regulator of sporulation, Spo0A, in low G+C Gram-positive endospore-forming bacteria.

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Surfaces of Spo0A and RNA Polymerase Sigma Factor A That Interact at the spoIIG Promoter in Bacillus subtilis

Abstract: In Bacillus subtilis, the DNA binding protein Spo0A activates transcription from two classes of promoters, those used by RNA polymerase containing the primary sigma factor, A (e.g., spoIIG), and those used by RNA polymerase containing the secondary sigma factor, H (e.g., spoIIA).

Cited by 13 publications

References 31 publications

Regulation of dev , an Operon That Includes Genes Essential for Myxococcus xanthus Development and CRISPR-Associated Genes and Repeats

Regulation of dev , an Operon That Includes Genes Essential for Myxococcus xanthus Development and CRISPR-Associated Genes and Repeats

Spo0A-Dependent Activation of an Extended −10 Region Promoter in Bacillus subtilis

Molecular insights into the initiation of sporulation in Gram-positive bacteria: new technologies for an old phenomenon

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