Role of sigma H in expression of the fumarase gene (citG) in vegetative cells of Bacillus subtilis 168

Price, V.A.; Feavers, Ian M.; Moir, Anne

doi:10.1128/jb.171.11.5933-5939.1989

Cited by 28 publications

(20 citation statements)

References 35 publications

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“…The concentration of UH increases about fivefold between the early stages of exponential growth and stationary phase in DS medium (J. Healy and R. Losick lated. These association constants have not been determined; however, this model is consistent with the observation that citGp2 is active in a spoOA mutant (16), in which there is a low concentration of CrH. Alternatively, the activity of these promoters could be regulated by different ancillary factors.…”

Section: Resultssupporting

confidence: 52%

“…Many of the genes transcribed by ErH are also transcribed by other polymerases. citG (7,16) and rpoD (4) are transcribed from promoters upstream from P2 and P3, respectively, that are probably used by ErA. The region downstream from pHI is read from a second upstream promoter at late stages of endospore formation (K. M. Tatti, unpublished results) as is the spolIA operon (5).…”

Section: Resultsmentioning

confidence: 99%

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Sigma H-directed transcription of citG in Bacillus subtilis

Tatti

Moir

et al. 1989

J Bacteriol

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The RNA polymerase sigma factor afH is essential for the onset of endospore formation in Bacillus subtilis.rH also is required for several additional stationary-phase-specific responses, including the normal expression of several genes that are required for the development of competence for DNA uptake. It is necessary to identify the genes that are transcribed by (rH RNA polymerase (EiH) in order to understand the role of this sigma factor during the transition from exponential growth to stationary phase. Feavers et al. (Mol. Gen. Genet. 211:465-471, 1988) proposed that MIG, the structural gene for fumarase, is transcribed from two promoters, one of which (citGp2 [P2]) may be used by EYH. It is likely that the citGp2 promoter is used by ErH because we found that this promoter was used accurately in vitro by EifH and directed expression of xylE in vivo. This xylE expression was dependent on spoOH, the structural gene for CfH, and was independent of the citGpl promoter. Comparison of the nucleotide sequences of several CfH-dependent promoters showed that these sequences were similar at two regions approximately 10 and 35 base pairs upstream from the start points of transcription. These sequences may signal recognition of these promoters by ECfH. Primer extension analyses were used to examine transcription from three fH-dependent promoters during growth and sporulation. The citGp2 promoter appeared to be active during the middle and late stages of exponential growth, whereas activation of the spoIIA promoter was delayed until after the end of exponential growth. Evidently, promoters used by EiH can display different temporal patterns of expression.Sigma H (uH) is one of at least nine different RNA polymerase sigma factors produced by the bacterium Bacillus subtilis (14, 15). Association of each sigma factor with the core subunits of RNA polymerase directs the holoenzyme to use a different class of transcriptional promoters. Since mutations in spoOH, the structural gene for aH (6), prevent the onset of sporulation, it is likely that RNA polymerase containing (7H (ECH) transcribes at least one gene that is necessary for the onset of sporulation. Mutations in spoOH also reduce the development of competence for DNA uptake (1) and the production of stationary-phase-specific extracellular enzymes (8). Evidently, EuH transcribes a diverse group of genes that play important roles in the bacterium's responses to nutrient depletion.The genes that are transcribed by EuH and necessary for the stationary-phase-specific responses are unknown. EuH has been shown to use two promoters, spoVG (22) and rpoDp3 (4), but transcription from these two promoters is not necessary for the onset of sporulation or the other stationary-phase-specific responses (18; R. H. Doi, personal communication). Moreover, recent results show that crA, the primary sigma factor in growing cells, directs the transcription of several sporulation-essential genes at the onset of endospore formation (9, 11; T. J. Kenney, K. York, P. Youngman, and C. P. Moran, J...

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Section: Resultssupporting

confidence: 52%

Section: Resultsmentioning

confidence: 99%

Sigma H-directed transcription of citG in Bacillus subtilis

Tatti

Moir

et al. 1989

J Bacteriol

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“…Sigma-H is required at an early stage of sporulation and directly activates transcription of several sporulation genes including spo0A, spo0F, kinA, spo0M, spoVG, and spoVS and the spoIIA operon (2,4,28,41,42,46,54,59,61). Sigma-H also directs the transcription of several members of the phr family of genes, which encode secreted peptide pheromones (31,37).…”

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confidence: 99%

Genome-Wide Analysis of the Stationary-Phase Sigma Factor (Sigma-H) Regulon ofBacillus subtilis

et al. 2002

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Sigma-H is an alternative RNA polymerase sigma factor that directs the transcription of many genes that function at the transition from exponential growth to stationary phase in Bacillus subtilis. Twenty-three promoters, which drive transcription of 33 genes, are known to be recognized by sigma-H-containing RNA polymerase. To identify additional genes under the control of sigma-H on a genome-wide basis, we carried out transcriptional profiling experiments using a DNA microarray containing >99% of the annotated B. subtilis open reading frames. In addition, we used a bioinformatics-based approach aimed at the identification of promoters recognized by RNA polymerase containing sigma-H. This combination of approaches was successful in confirming most of the previously described sigma-H-controlled genes. In addition, we identified 26 putative promoters that drive expression of 54 genes not previously known to be under the direct control of sigma-H. Based on the known or inferred function of most of these genes, we conclude that, in addition to its previously known roles in sporulation and competence, sigma-H controls genes involved in many physiological processes associated with the transition to stationary phase, including cytochrome biogenesis, generation of potential nutrient sources, transport, and cell wall metabolism.Bacterial sigma factors are positive regulators of gene expression that interact with core RNA polymerase and direct the initiation of transcription from defined promoter sequences (22, 25). The major sigma factor in most bacteria, sigma-A, is required for expression of many of the so-called housekeeping functions and the bulk of the RNA during growth. Many bacteria have multiple alternative sigma factors, which are responsible for directing transcription of specialized gene sets. Bacillus subtilis has at least 17 alternative sigma factors which are involved in a variety of processes, including certain stress responses, chemotaxis, and motility (25,30). One of the more dramatic examples of gene regulation by alternative sigma factors is the process of endospore formation (sporulation) in B. subtilis. The sporulation program of gene expression in B. subtilis is carried out under the direction of five alternative sigma factors whose activities are subject to spatial and temporal control (14, 51). Here we report the results of transcriptional profiling experiments aimed at identifying, on a genome-wide basis, genes under the control of one of these sigma factors, sigma-H.Sigma-H, the sigH (spo0H) gene product, directs the transcription of several genes that function in the transition from exponential growth to stationary phase, including the initiation of spore formation and entry into the state of genetic competence (1,7,12,20). Sigma-H is required at an early stage of sporulation and directly activates transcription of several sporulation genes including spo0A, spo0F, kinA, spo0M, spoVG, and spoVS and the spoIIA operon (2,4,28,41,42,46,54,59,61). Sigma-H also directs the transcription of several mem...

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“…4 transcription assays. The preparation of RNA polymerase that was used has been shown to catalyze transcription from the spoVG promoter and the citG P2 promoter (25,29). If the UAS affects spoVG transcription by enhancing the affinity of RNA polymerase for the promoter without the aid of any additional factors, such as a positive regulatory protein, then the reduction in promoter activity caused by the spoVGD, spoVGJB, and spoVG210 mutations should be observed in vitro by using purified EcrH.…”

Section: Resultsmentioning

confidence: 99%

Analysis of the upstream activating sequence and site of carbon and nitrogen source repression in the promoter of an early-induced sporulation gene of Bacillus subtilis

Frisby

Zuber

1991

J Bacteriol

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The transcription from the spoVG promoter of Bacillus subtilis is induced at the start of the stationary phase of growth and is dependent on the expression of the spoOA, spoOB, and spoOH genes. It is repressed in cells grown in the presence of excess glucose and glutamine and is under the negative control of the abrB gene. The spoOA and spoOB gene products function to suppress the negative control exerted by abrB. Transcription initiation requires the form of RNA polymerase holoenzyme that contains the spoOH gene product, cH. Optimal transcription also requires an upstream A-T-rich region termed the upstream activating sequence (UAS). The mechanism of UAS function was examined through mutational analysis of the spoVG promoter region. Deletion of the UAS or positioning the UAS one half turn or one full turn of the DNA helix upstream of its location in wild-type spoVG resulted in a severe reduction in promoter activity. Deletion of most of the UAS abolished the abrB-dependent repression of spoVG transcription. Higher activity was observed when the UAS was inserted 10 bp (one turn of the helix) upstream than when the sequence was repositioned either 5 or 13 bp upstream. Sequences upstream of the UAS were found not to be involved with the position-dependent function of the UAS. Positioning the UAS 42 or 116 bp upstream eliminated the stimulatory effect of the sequence on spoVG transcription. These data indicate that the UAS functions effectively when it is in close proximity to the -35 region. In vitro transcription analysis indicated that the deletion and insertion mutations affecting the UAS impair RNA polymerase-spoVG promoter interaction. Deletion of the UAS showed that the negative effect of exogenous glucose and glutamine is not dependent on the UAS but is exerted at a site within or near the -35 and -10 regions.Endospore formation in Bacillus subtilis is a developmental process triggered by nutritional deprivation. It can be divided into seven well-defined stages, beginning with the initiation of sporulation (stage 0) and ending with mother cell lysis (stage VII) and liberation of the mature spore (24). Initiation of sporulation requires the products of at least seven spoO genes (spoOA, spoOB, spoOE, spoOF, spoOH, spoOJ, and spoOK) to begin the cascade of events leading to the expression of numerous sporulation-associated genes (13,24). A mutation in any one of the spoO genes blocks the sporulation process before any morphological changes within the cell are observed. The spoVG gene has served as a system for examining the activation of gene transcription at the onset of sporulation because it is under spoO control and is induced as cells enter the stationary phase of growth in sporulation medium (23,33). Furthermore, transcription of spoVG is repressed when cells are grown in a medium containing a high concentration of glucose and glutamine, preferred carbon and nitrogen sources which inhibit sporulation when present in excess (24,27).Transcription of spoVG is dependent upon the products of spoOA, spoOB, a...

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Role of sigma H in expression of the fumarase gene (citG) in vegetative cells of Bacillus subtilis 168

Cited by 28 publications

References 35 publications

Sigma H-directed transcription of citG in Bacillus subtilis

Sigma H-directed transcription of citG in Bacillus subtilis

Genome-Wide Analysis of the Stationary-Phase Sigma Factor (Sigma-H) Regulon ofBacillus subtilis

Analysis of the upstream activating sequence and site of carbon and nitrogen source repression in the promoter of an early-induced sporulation gene of Bacillus subtilis

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