Sigma H-directed transcription of citG in Bacillus subtilis

Tatti, Kathleen M.; rd, H L Carter; Moir, Anne; Moran, Charles P.

doi:10.1128/jb.171.11.5928-5932.1989

Cited by 52 publications

(40 citation statements)

References 23 publications

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“…4). The P2-directed activity was dependent on an intact spoOH gene, as is discussed below and in the accompanying paper (32). The suppression of fumarase activity by readily metabolizable carbon sources is therefore exerted at the level of citGp2 transcription.…”

Section: Methodsmentioning

confidence: 82%

“…Transcription from citGp2 is dependent on a functional spoOH gene, with the consequence that spoOH is required for efficient fumarase expression (9). spoOH encodes an alternative sigma factor (sigma H), which together with the RNA polymerase core enzyme directs transcription from citGp2 (32). Thus, the analysis of the transcriptional control of fumarase in B. subtilis is complicated by the presence of multiple promoters and the involvement of differentially regulated multiple holoenzyme forms of RNA polymerase.…”

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confidence: 99%

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

1989

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The fumarase gene (citG) of Bacillus subtilis is transcribed from two promoter regions, cilGpl and citGp2 (P1 and P2); the P2 promoter is used by the EUH form of RNA polymerase. In order to study the role of P1 and P2 in citG expression, the promoter region and various deletion derivatives that effectively separate P1 and P2 were fused to the Escherichia coli ,-galactosidase gene (lacZ) and introduced into the chromosome in single copy at the amyE locus. P1 functioned to provide a relatively low and stable basal level of fumarase activity throughout growth. In contrast, P2 activity was found to vary over at least a 50-fold range and was responsible for regulating fumarase activity during growth and sporulation in a rich medium and in response to changes in carbon source. To further investigate the role of sigma H in fumarase regulation, citGp2-lacZ fusions were introduced into a strain in which the expression of the chromosomal spoOH gene was under the control of the isopropylthiogalactopyranoside-inducible spac promoter. Induction of p,P did not lead to P2 induction, suggesting that cilG expression is not regulated at the level of spoOH transcription.The tricarboxylic acid (TCA) cycle in Bacillus subtilis plays a dual role in metabolism, providing both energy (when glycolysis is unable to fulfill the cell's energy requirements) and biosynthetic intermediates. The cycle can be regarded as two pathways: the tricarboxylic pathway (citrate to 2-ketoglutarate) and the dicarboxylic pathway (succinyl coenzyme A to oxaloacetate). These pathways are subject to independent but overlapping regulation (15,27). Levels of fumarase, a component of the dicarboxylic acid pathway, increase during vegetative growth in a rich medium to peak between 1 and 2 h after the end of exponential growth (9). Levels are highest in a lactate minimal medium (when both anabolic and energy-generating functions of the cycle are of prime importance) and are depressed by rapidly metabolizable carbon sources such as glucose and casamino acids (15).The fumarase gene (citG) has been cloned and sequenced (22,24,25), and it has been shown to be transcribed from two promoter regions, citGpl (P1) and citGp2 (P2); citGpl appears to be a complex of two overlapping promoters, Pla and Plb (9). citGpl and citGp2 are separated by a short open reading frame (orfA), which has the potential to encode a small (61-residue) polypeptide. Transcription from citGp2 is dependent on a functional spoOH gene, with the consequence that spoOH is required for efficient fumarase expression (9). spoOH encodes an alternative sigma factor (sigma H), which together with the RNA polymerase core enzyme directs transcription from citGp2 (32). Thus, the analysis of the transcriptional control of fumarase in B. subtilis is complicated by the presence of multiple promoters and the involvement of differentially regulated multiple holoenzyme forms of RNA polymerase.To tackle this problem, fusions of separated citGpl and citGp2 promoters to the Escherichia coli lacZ gene were constructed, s...

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Section: Methodsmentioning

confidence: 82%

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confidence: 99%

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

1989

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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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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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“…Recently, a connection between the heat shock response and the initiation of solvent formation in C. acetobutylicum was proposed (20,25). In the regulation of the heat shock response of E. coli (23) and the sporulation process of B. subtilis (24), alternate sigma factors play an important role. It will be of interest to know whether the RNA polymerase from C. acetobutylicum cells, which switch from acid to solvent production, initiate sporulation, and/or are exposed to other forms of stress, contains different sigma factors controlling the expression of respective genes.…”

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Purification and characterization of the DNA-dependent RNA polymerase from Clostridium acetobutylicum

Pich

Bahl

1991

J Bacteriol

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The DNA-dependent RNA polymerase (EC 2.7.7.6) from Clostridium acetobutylicum DSM 1731 has been purified to homogeneity and characterized. The purified enzyme was composed of four subunits and had a molecular mass of 370,000 Da. Western immunoblot analysis with polyclonal antibodies against the sigma 70 subunit of Escherichia coli RNA polymerase identified the 46,000-Da subunit as an immunologically and probably functionally related protein. The other three subunits of 128,000, 117,000, and 42,000 Da are tentatively analogous to the beta, beta', and alpha subunits, respectively, of other eubacterial RNA polymerases. The RNA polymerase activity was completely dependent on Mg2+, nucleoside triphosphates, and a DNA template. The presence of Mg2+ or Mn2+ in buffers used for purification or storage caused irreversible inactivation of the RNA polymerase.

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Sigma H-directed transcription of citG in Bacillus subtilis

Cited by 52 publications

References 23 publications

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

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

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

Purification and characterization of the DNA-dependent RNA polymerase from Clostridium acetobutylicum

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