Temporal and selective association of multiple sigma factors with RNA polymerase during sporulation in Bacillus subtilis

Fujita, Masaya

doi:10.1046/j.1365-2443.2000.00307.x

Cited by 69 publications

(66 citation statements)

References 24 publications

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“…Polyclonal anti-Spo0A antibodies (Fujita 2000) were used for the detection of Spo0A and Spo0A*. Polyclonal GFP antibodies (Rudner and Losick 2002) were used for the detection of GFP-tagged KinA.…”

Section: Immunoblot Analysismentioning

confidence: 99%

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Evidence that entry into sporulation in Bacillus subtilis is governed by a gradual increase in the level and activity of the master regulator Spo0A

Fujita¹,

Losick²

2005

Genes Dev.

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267

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The transcription factor Spo0A is a master regulator for entry into sporulation in the bacterium Bacillus subtilis, but it has been uncertain whether activation of Spo0A is sufficient to trigger development. Spo0A, a member of the response regulator family of gene-control proteins, is activated by phosphorylation via a multicomponent phosphorelay in response to conditions of nutrient limitation. We now report that sporulation can be triggered with high efficiency in cells in the exponential phase of growth in rich medium by artificial induction of the synthesis of any one of three histidine kinases that feed phosphoryl groups into the relay. We further show that the levels of Spo0A protein and activity increase gradually over the first 2 h of sporulation both under conditions of nutrient limitation and in response to induction of kinase synthesis. Evidence indicates that this gradual increase in Spo0A protein and activity plays a critical role in triggering sporulation and requires the action of the phosphorelay.[Keywords: Bacillus subtilis; sporulation; development; response regulator; transcription; phosphorelay] Supplemental material is available at http://www.genesdev.org.

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Section: Immunoblot Analysismentioning

confidence: 99%

“…Polyclonal GFP antibodies (Rudner and Losick 2002) were used for the detection of GFP-tagged KinA. Polyclonal A antibodies (Fujita 2000) were used for the detection of A .…”

Section: Immunoblot Analysismentioning

confidence: 99%

Evidence that entry into sporulation in Bacillus subtilis is governed by a gradual increase in the level and activity of the master regulator Spo0A

Fujita¹,

Losick²

2005

Genes Dev.

Self Cite

267

320

View full text Add to dashboard Cite

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“…Recent studies have shown that core RNA polymerase and s A levels remain constant during the transition from vegetative growth to sporulation (Fujita, 2000), and several lines of evidence suggest that s A -directed transcription takes place and is important even at late stages of sporulation (Fujita, 2000;Fukushima et al, 2003;Li & Piggot, 2001;Ohashi et al, 2003). A metabolic-pathwayoriented inspection of the temporal expression pattern revealed that certain classes of housekeeping genes continued to be expressed at a high level during sporulation, at least until 150 min after the initiation of sporulation.…”

Section: Differentiation Of the Compartment-specific Sporulation Regumentioning

confidence: 99%

Genome-wide analysis of temporally regulated and compartment-specific gene expression in sporulating cells of Bacillus subtilis

et al. 2005

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Temporal and compartment-specific control of gene expression during sporulation in Bacillus subtilis is governed by a cascade of four RNA polymerase subunits. s F in the prespore and s E in the mother cell control early stages of development, and are replaced at later stages by s G and s K , respectively. Ultimately, a comprehensive description of the molecular mechanisms underlying spore morphogenesis requires the knowledge of all the intervening genes and their assignment to specific regulons. Here, in an extension of earlier work, DNA macroarrays have been used, and members of the four compartment-specific sporulation regulons have been identified. Genes were identified and grouped based on: i) their temporal expression profile and ii) the use of mutants for each of the four sigma factors and a bofA allele, which allows s K activation in the absence of s G . As a further test, artificial production of active alleles of the sigma factors in non-sporulating cells was employed. A total of 439 genes were found, including previously characterized genes whose transcription is induced during sporulation: 55 in the s F regulon, 154 s E -governed genes, 113 s G -dependent genes, and 132 genes under s K control. The results strengthen the view that the activities of s F , s E , s G and s K are largely compartmentalized, both temporally as well as spatially, and that the major vegetative sigma factor (s A ) is active throughout sporulation. The results provide a dynamic picture of the changes in the overall pattern of gene expression in the two compartments of the sporulating cell, and offer insight into the roles of the prespore and the mother cell at different times of spore morphogenesis.

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“…The rabbit polyclonal antibodies that bind to A and Spo0A (Fujita 2000) were used at 1:10,000 and 1:5000 dilutions, respectively.…”

Section: Immunofluorescence Microscopymentioning

confidence: 99%

The master regulator for entry into sporulation inBacillus subtilisbecomes a cell-specific transcription factor after asymmetric division

Fujita¹,

Losick²

2003

Genes Dev.

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108

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Gene transcription at the onset of sporulation in Bacillus subtilis is governed by Spo0A, a member of the response regulator family of transcription factors. Spo0A is traditionally viewed as the master regulator for entry into development. We now report that Spo0A continues to function after the initiation phase of sporulation and that it becomes a cell-specific transcription factor when the sporangium is divided into a mother cell and forespore. We observed that (1) Spo0A and Spo0A-directed gene transcription reached high levels in the mother cell; (2) an activated form of Spo0A impaired sporulation when produced in the forespore but not when produced in the mother cell; and (3) an inhibitor of Spo0A called Spo0A-N impaired sporulation and Spo0A-directed transcription when produced in the mother cell but not when produced in the forespore. Spo0A-N, which corresponds to the NH 2 -terminal domain of Spo0A, was shown to compete with the full-length response regulator for phosphorylation by the phosphorelay protein Spo0B. We propose that Spo0A is the earliest-acting transcription factor in the mother-cell line of gene expression and that in terms of abundance and transcriptional activity Spo0A may function predominantly as a cell-specific regulatory protein.[Keywords: Bacillus subtilis; sporulation; response regulator; transcription; asymmetry] Supplemental material is available at http://www.genesdev.org.

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Temporal and selective association of multiple sigma factors with RNA polymerase during sporulation in Bacillus subtilis

Cited by 69 publications

References 24 publications

Evidence that entry into sporulation in Bacillus subtilis is governed by a gradual increase in the level and activity of the master regulator Spo0A

Evidence that entry into sporulation in Bacillus subtilis is governed by a gradual increase in the level and activity of the master regulator Spo0A

Genome-wide analysis of temporally regulated and compartment-specific gene expression in sporulating cells of Bacillus subtilis

The master regulator for entry into sporulation inBacillus subtilisbecomes a cell-specific transcription factor after asymmetric division

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