scholarly journals Role of the Anti-Sigma Factor SpoIIAB in Regulation of σG during Bacillus subtilis Sporulation

2004 ◽  
Vol 186 (12) ◽  
pp. 4000-4013 ◽  
Author(s):  
Mónica Serrano ◽  
Alexandre Neves ◽  
Cláudio M. Soares ◽  
Charles P. Moran ◽  
Adriano O. Henriques
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Mother Cell ◽  
In Vivo Studies ◽  
Specific Gene ◽  
Rapid Induction ◽  
Inactive Form ◽  
Entire Cell

ABSTRACT RNA polymerase sigma factor σF initiates the prespore-specific program of gene expression during Bacillus subtilis sporulation. σF governs transcription of spoIIIG, encoding the late prespore-specific regulator σG. However, transcription of spoIIIG is delayed relative to other genes under the control of σF, and after synthesis, σG is initially kept in an inactive form. Activation of σG requires the complete engulfment of the prespore by the mother cell and expression of the spoIIIA and spoIIIJ loci. We screened for random mutations in spoIIIG that bypassed the requirement for spoIIIA for the activation of σG. We found a mutation (spoIIIGE156K) that resulted in an amino acid substitution at position 156, which is adjacent to the position of a mutation (E155K) previously shown to prevent interaction of SpoIIAB with σG. Comparative modelling techniques and in vivo studies suggested that the spoIIIGE156K mutation interferes with the interaction of SpoIIAB with σG. The σGE156K isoform restored σG-directed gene expression to spoIIIA mutant cells. However, expression of sspE-lacZ in the spoIIIA spoIIIGE156K double mutant was delayed relative to completion of the engulfment process and was not confined to the prespore. Rather, β-galactosidase accumulated throughout the entire cell at late times in development. This suggests that the activity of σGE156K is still regulated in the prespore of a spoIIIA mutant, but not by SpoIIAB. In agreement with this suggestion, we also found that expression of spoIIIGE156K from the promoter for the early prespore-specific gene spoIIQ still resulted in sspE-lacZ induction at the normal time during sporulation, coincidently with completion of the engulfment process. In contrast, transcription of spoIIIGE156K, but not of the wild-type spoIIIG gene, from the mother cell-specific spoIID promoter permitted the rapid induction of sspE-lacZ expression. Together, the results suggest that SpoIIAB is either redundant or has no role in the regulation of σG in the prespore.

scholarly journals Expression of spoIIIJ in the Prespore Is Sufficient for Activation of σG and for Sporulation in Bacillus subtilis

2003 ◽  
Vol 185 (13) ◽  
pp. 3905-3917 ◽  
Author(s):  
Mónica Serrano ◽  
Luísa Côrte ◽  
Jason Opdyke ◽  
Charles P. Moran, ◽  
Adriano O. Henriques
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Rna Polymerase ◽  
Sigma Factor ◽  
Vegetative Growth ◽  
Mother Cell ◽  
Asymmetric Division ◽  
Developmental Program ◽  
Inactive Form

ABSTRACT During sporulation in Bacillus subtilis, the prespore-specific developmental program is initiated soon after asymmetric division of the sporangium by the compartment-specific activation of RNA polymerase sigma factor σF. σF directs transcription of spoIIIG, encoding the late forespore-specific regulator σG. Following synthesis, σG is initially kept in an inactive form, presumably because it is bound to the SpoIIAB anti-sigma factor. Activation of σG occurs only after the complete engulfment of the prespore by the mother cell. Mutations in spoIIIJ arrest sporulation soon after conclusion of the engulfment process and prevent activation of σG. Here we show that σG accumulates but is mostly inactive in a spoIIIJ mutant. We also show that expression of the spoIIIGE155K allele, encoding a form of σG that is not efficiently bound by SpoIIAB in vitro, restores σG-directed gene expression to a spoIIIJ mutant. Expression of spoIIIJ occurs during vegetative growth. However, we show that expression of spoIIIJ in the prespore is sufficient for σG activation and for sporulation. Mutations in the mother cell-specific spoIIIA locus are known to arrest sporulation just after completion of the engulfment process. Previous work has also shown that σG accumulates in an inactive form in spoIIIA mutants and that the need for spoIIIA expression for σG activation can be circumvented by the spoIIIGE155K allele. However, in contrast to the case for spoIIIJ, we show that expression of spoIIIA in the prespore does not support efficient sporulation. The results suggest that the activation of σG at the end of the engulfment process involves the action of spoIIIA from the mother cell and of spoIIIJ from the prespore.

scholarly journals Establishment of Prespore-Specific Gene Expression inBacillus subtilis: Localization of SpoIIE Phosphatase and Initiation of Compartment-Specific Proteolysis

1998 ◽  
Vol 180 (13) ◽  
pp. 3276-3284 ◽  
Author(s):  
Peter J. Lewis ◽  
Ling Juan Wu ◽  
Jeffery Errington
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Mother Cell ◽  
Immunofluorescence Microscopy ◽  
The Other ◽  
Specific Gene ◽  
Specific Gene Expression

ABSTRACT Immunofluorescence microscopy was used to study the establishment of compartment-specific transcription during sporulation inBacillus subtilis. Analysis of the distribution of the anti-anti-sigma factor, SpoIIAA, in a variety of mutant backgrounds supports a model in which the SpoIIE phosphatase, which activates SpoIIAA by dephosphorylation, is sequestered onto the prespore face of the asymmetric septum. Thus, prespore-specific gene expression apparently arises as a result of the compartmentalization of SpoIIE protein. The results also suggest the existence of at least two compartment-specific programs of proteolysis, one dependent on the mother cell-specific sigma factor ςE and the other dependent on the prespore-specific sigma factor ςF.

scholarly journals A Dispensable Role for Forespore-Specific Gene Expression in Engulfment of the Forespore during Sporulation ofBacillus subtilis

2000 ◽  
Vol 182 (10) ◽  
pp. 2919-2927 ◽  
Author(s):  
Ya-Lin Sun ◽  
Marc D. Sharp ◽  
Kit Pogliano
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Sigma Factor ◽  
Mother Cell ◽  
Spore Production ◽  
Specific Gene ◽  
Ofbacillus Subtilis ◽  
Formation Pathway ◽  
Specific Gene Expression

ABSTRACT During the stage of engulfment in the Bacillus subtilisspore formation pathway, the larger mother cell engulfs the smaller forespore. We have tested the role of forespore-specific gene expression in engulfment using two separate approaches. First, using an assay that unambiguously detects sporangia that have completed engulfment, we found that a mutant lacking the only forespore-expressed engulfment protein identified thus far, SpoIIQ, is able to efficiently complete engulfment under certain sporulation conditions. However, we have found that the mutant is defective, under all conditions, in the expression of the late-forespore-specific transcription factor ςG; thus, SpoIIQ is essential for spore production. Second, to determine if engulfment could proceed in the absence of forespore-specific gene expression, we made use of a strain in which activation of the mother cell-specific sigma factor ςE was uncoupled from forespore-specific gene expression. Remarkably, engulfment occurred in the complete absence of ςF-directed gene expression under the same conditions permissive for engulfment in the absence of SpoIIQ. Our results demonstrate that forespore-specific gene expression is not essential for engulfment, suggesting that the machinery used to move the membranes around the forespore is within the mother cell.

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

Microbiology ◽  
2005 ◽  
Vol 151 (2) ◽  
pp. 399-420 ◽  
Author(s):  
Leif Steil ◽  
Mónica Serrano ◽  
Adriano O. Henriques ◽  
Uwe Völker
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Mother Cell ◽  
Molecular Mechanisms ◽  
Sigma Factors ◽  
Specific Gene ◽  
Temporal Expression ◽  
Control Of Gene Expression ◽  
Genome Wide ◽  
Specific Control

Temporal and compartment-specific control of gene expression during sporulation in Bacillus subtilis is governed by a cascade of four RNA polymerase subunits. σ F in the prespore and σ E in the mother cell control early stages of development, and are replaced at later stages by σ G and σ 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 σ K activation in the absence of σ 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 σ F regulon, 154 σ E-governed genes, 113 σ G-dependent genes, and 132 genes under σ K control. The results strengthen the view that the activities of σ F, σ E, σ G and σ K are largely compartmentalized, both temporally as well as spatially, and that the major vegetative sigma factor (σ 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.

scholarly journals Dual-Specificity Anti-sigma Factor Reinforces Control of Cell-Type Specific Gene Expression in Bacillus subtilis

PLoS Genetics ◽  
2015 ◽  
Vol 11 (4) ◽  
pp. e1005104 ◽  
Author(s):  
Mónica Serrano ◽  
JinXin Gao ◽  
João Bota ◽  
Ashley R. Bate ◽  
Jeffrey Meisner ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Specific Gene ◽  
Cell Type ◽  
Specific Gene Expression ◽  
Dual Specificity ◽  
Cell Type Specific

scholarly journals Developmental Gene Expression in Bacillus subtilis crsA47 Mutants Reveals Glucose-Activated Control of the Gene for the Minor Sigma Factor ςH

2001 ◽  
Vol 183 (16) ◽  
pp. 4814-4822 ◽  
Author(s):  
Laurie G. Dixon ◽  
Steve Seredick ◽  
Martin Richer ◽  
George B. Spiegelman
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Glucose Repression ◽  
Gene Promoter ◽  
Growth Media ◽  
Regulatory Constraints ◽  
Expression Of Genes

ABSTRACT The presence of excess glucose in growth media prevents normal sporulation of Bacillus subtilis. The crsA47mutation, located in the gene for the vegetative phase sigma factor (ςA) results in a glucose-resistant sporulation phenotype. As part of a study of the mechanisms whereby the mutation in ςA overcomes glucose repression of sporulation, we examined the expression of genes involved in sporulation initiation in the crsA47 background. The crsA47 mutation had a significant impact on a variety of genes. Changes to stage II gene expression could be linked to alterations in the expression of thesinI and sinR genes. In addition, there was a dramatic increase in the expression of genes dependent on the minor sigma factor ςH. This latter change was paralleled by the pattern of spo0H gene transcription in cells with thecrsA47 mutation. In vitro analysis of RNA polymerase containing ςA47 indicated that it did not have unusually high affinity for the spo0H gene promoter. The in vivo pattern of spo0H expression is not predicted by the known regulatory constraints on spo0H and suggests novel regulation mechanisms that are revealed in the crsA47background.

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