Analysis of transcriptional control of the gerD spore germination gene of Bacillus subtilis 168

Kemp, E. Helen; Sammons, Rachel; Moir, Anne; Sun, Dongxu; Setlow, Peter

doi:10.1128/jb.173.15.4646-4652.1991

Cited by 61 publications

(44 citation statements)

References 26 publications

(64 reference statements)

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“…The apparent increase in sspBlacZ expression is presumably due to a greater efficiency of breakage of the forespores during assay of the spoVB mutant. Since this result contradicts earlier reports of severely decreased forespore-specific gene expression in the spoIIIF590 mutant (9,16,17,24), the spoIIIF590 and spoVB9J mutations were introduced by congression in the JH642 background. Expression of the sspB-1acZ fusion in these two strains was found to be similar to the expression observed in the spoVB MLS' mutant (data not shown).…”

Section: Resultscontrasting

confidence: 55%

“…The close proximity of the two loci was confirmed by the isolation of a recombinant transducing phage able to complement both mutations (8). Interestingly, the spoIIIFS90 mutation has been reported to strongly decrease transcription of spollIG, the gene encoding the forespore-specific uJ factor, uG (16), as well as expression of several forespore-expressed genes (9,17,24). We show here that spoVB9J and spoIIIFS90 are alleles of the same gene and that characteristics previously ascribed to spoIIIF590 (sporulation blockage at stage III and decrease of foresporespecific gene expression) are probably due to a secondary mutation present in some strains.…”

mentioning

confidence: 89%

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Cloning, characterization, and expression of the spoVB gene of Bacillus subtilis

Popham

Stragier

1991

J Bacteriol

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Mutation of the spoVB gene in Bacillus subtilis causes the production of spores containing a defective cortex and unable to acquire heat resistance. The spoVB locus is highly linked to another spo locus, spoIIIF, characterized by a single mutation (I. L. Lamont and J. Mandelstam, J. Gen. Microbiol. 130:1253-1261. A 18-kb DNA region overlapping the spoIIIF-spoVB region was cloned in successive steps starting from a Tn917 insertion in the nic locus. The exact location of the spollIF and spoVB loci was defined with various integrative plasmids carrying subfragments of that region. DNA sequencing established that spollIF and spoVB are a single monocistronic locus encoding a 518-amino-acid polypeptide with features of an integral membrane protein. The precise location of the spoIIIF590 and spoVB91 mutations in that unique open reading frame was determined, and both mutations were sequenced. A null mutation was engineered in the spoIIIF-spoVB locus and led to a typical spoVB phenotype, identical to the phenotype created by either spoIIIF590 or spoVB9l, suggesting that the original spollIF mutant contained a secondary mutation arresting sporulation at an earlier stage. A transcriptional spoVB-lacZ fusion was constructed, and its expression was found to be directly dependent on RNA polymerase containing SE. A null mutation of spoVB had no effect on expression of sspB and cotA, members of the 7G-and rK-controlled regulons respectively, while expression of cotC, a member of the latest known mother cell regulon, was delayed and strongly reduced. These results are consistent with SpoVB being involved in cortex biosynthesis and affecting only indirectly expression of late sporulation genes.Sporulation in bacilli involves a succession of coordinated steps in which two cellular compartments within a sporangium supply different components to the spore (23). The smaller compartment, the forespore, produces the smaller DNA-binding proteins which confer UV resistance to the spore (25, 33) and apparently the inner peptidoglycan layer surrounding the spore, the germ cell wall (38). The larger compartment, the mother cell, provides the coat proteins which form the outermost layers of the spore (3) and some activities required for synthesis of the cortex (38), a thick outer layer of peptidoglycan. The cortex seems to be involved in attaining or maintaining the dehydrated state of the spore protoplasm, a condition required for heat resistance (12), while the coat proteins provide a barrier to agents such as lysozyme (15,43). The successive production of these spore components is programmed by a series of regulatory genes which appear to activate their respective steps in response to morphological signals of completion of the preceding steps (36).A large number of mutations which block the sporulation process at various morphological steps have been isolated in Bacillus subtilis (28). Analysis of these mutations has shown that many of them are in regulatory genes, presumably because mutation of a factor which has pleiotropic effects on ...

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Section: Resultscontrasting

confidence: 55%

mentioning

confidence: 89%

Cloning, characterization, and expression of the spoVB gene of Bacillus subtilis

Popham

Stragier

1991

J Bacteriol

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“…This fragment contains the coding sequence for the first 108 amino acids of CWLA and 672 bp of upstream sequence (Foster, 1991). After ligation into SmaI-cut and dephosphorylated pAZlO6 (Kemp et al, 1991 ;A. Zuberi & R. Doi, unpublished data) this construct was used to transform E. coli DH5a (Hanahan, 1983).…”

Section: Methodsmentioning

confidence: 99%

Analysis of Bacillus subtilis 168 prophage-associated lytic enzymes; identification and characterization of CWLA-related prophage proteins

Foster

1993

Journal of General Microbiology

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The CWLA lytic amidase of BaciZZus subtilis 168 was purified and antisera raised against the purified protein.No expression of CWZA could be demonstrated under any conditions by the use of the antisera and cwZA::ZacZ fusion analysis. Two lytic enzymes of apparent molecular masses 34 and 30 kDa (as measured by renaturing SDS-PAGE) were found to be mitomycin C-inducible, the larger of which corresponds to a protein immunologically related to CWLA. Both of these inducible lysins were found to be encoded by prophage PBSX. Prophage SPP was shown by renaturing SDS-PAGE to produce a 43 kDa lytic enzyme unrelated immunologically to CWLA. The smaller of the two PBSX enzymes was purified and found to be an N-acetylmuramyl-L-alanine amidase of 32 kDa (as measured by SDS-PAGE and Coomassie blue staining) which cross-reacts only weakly with the anti-CWLA sera. The potential origin of CWZA and its possible relationship to the other phage lytic enzymes are discussed.

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“…4a). Several genes in the s F regulon have functions in spore protection or spore germination: the katX-encoded catalase, for example, is implicated in spore protection against hydrogen peroxide (Bagyan et al, 1998); the mutTA gene encodes an antimutator 8-oxodGTPase (Ramirez et al, 2004); the sspN and tlp genes, which code for small acid-soluble spore proteins (SASP), act by shielding the prespore chromosome (CabreraHernandez et al, 1999); gerD is required for efficient germination in response to L-alanine and to a mixture of glucose, fructose, L-asparagine and KCl (Kemp et al, 1991); and gpr encodes a protease involved in SASP protein degradation during spore germination (Sussman & Setlow, 1991). Interestingly, our analysis identified the gene (yyaC) for a second possible GPR-like protease in the s F regulon (Fig.…”

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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Analysis of transcriptional control of the gerD spore germination gene of Bacillus subtilis 168

Cited by 61 publications

References 26 publications

Cloning, characterization, and expression of the spoVB gene of Bacillus subtilis

Cloning, characterization, and expression of the spoVB gene of Bacillus subtilis

Analysis of Bacillus subtilis 168 prophage-associated lytic enzymes; identification and characterization of CWLA-related prophage proteins

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

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