scholarly journals Regulation and Characterization of a Newly Deduced Cell Wall Hydrolase Gene (cwlJ) Which Affects Germination of Bacillus subtilis Spores

1998 ◽  
Vol 180 (6) ◽  
pp. 1375-1380 ◽  
Author(s):  
Shu Ishikawa ◽  
Kunio Yamane ◽  
Junichi Sekiguchi
Keyword(s):  
Bacillus Subtilis ◽  
Type Strain ◽  
Wild Type Strain ◽  
Catalytic Domain ◽  
Slow Release ◽  
Dipicolinic Acid ◽  
Northern Hybridization ◽  
Predicted Amino Acid Sequence ◽  
Wild Type

ABSTRACT The predicted amino acid sequence of Bacillus subtilis ycbQ (renamed cwlJ) exhibits high similarity to those of the deduced C-terminal catalytic domain of SleBs, the specific cortex-hydrolyzing enzyme of B. cereus and the deduced one of B. subtilis. We constructed acwlJ::lacZ fusion in the B. subtilischromosome. The β-galactosidase activity and results of Northern hybridization and primer extension analyses of the cwlJgene indicated that it is transcribed by EςE RNA polymerase. cwlJ-deficient spores responded to bothl-alanine and AGFK, the A 580 values of spore suspensions decreased more slowly than in the case of the wild-type strain, and the mutant spores released less dipicolinic acid than did those of the wild-type strain during germination. However, the mutant spores released only slightly less hexosamine than did the wild-type spores. In contrast, B. subtilis sleB spores did not release hexosamine at a significant level. While cwlJand sleB spores were able to germinate, CJSB (cwlJ sleB) spores could not germinate but exhibited initial germination reactions, e.g., partial decrease inA 580 and slow release of dipicolinic acid. CJSB spores became slightly gray after 6 h in the germinant, but their refractility was much greater than that of sleB mutant spores. The roles of the sleB and cwlJmutations in germination and spore maturation are also discussed.

scholarly journals A Polysaccharide Deacetylase Gene (pdaA) Is Required for Germination and for Production of Muramic δ-Lactam Residues in the Spore Cortex of Bacillus subtilis

2002 ◽  
Vol 184 (21) ◽  
pp. 6007-6015 ◽  
Author(s):  
Tatsuya Fukushima ◽  
Hiroki Yamamoto ◽  
Abdelmadjid Atrih ◽  
Simon J. Foster ◽  
Junichi Sekiguchi
Keyword(s):  
Bacillus Subtilis ◽  
Phase Contrast ◽  
Biosynthetic Pathway ◽  
Wild Type Strain ◽  
Dipicolinic Acid ◽  
Northern Hybridization ◽  
Predicted Amino Acid Sequence ◽  
Wild Type ◽  
Contrast Microscopy ◽  
Acid Release

ABSTRACT The predicted amino acid sequence of Bacillus subtilis yfjS (renamed pdaA) exhibits high similarity to those of several polysaccharide deacetylases. β-Galactosidase fusion experiments and results of Northern hybridization with sporulation sigma mutants indicated that the pdaA gene is transcribed by EσG RNA polymerase. pdaA-deficient spores were bright by phase-contrast microscopy, and the spores were induced to germination on the addition of l-alanine. Germination-associated spore darkening, a slow and partial decrease in absorbance, and slightly lower dipicolinic acid release compared with that by the wild-type strain were observed. In particular, the release of hexosamine-containing materials was lacking in the pdaA mutant. Muropeptide analysis indicated that the pdaA-deficient spores completely lacked muramic δ-lactam. A pdaA-gfp fusion protein constructed in strain 168 and pdaA-deficient strains indicated that the protein is localized in B. subtilis spores. The biosynthetic pathway of muramic δ-lactam is discussed.

scholarly journals Transcription of the Bacillus subtilis gerK Operon, Which Encodes a Spore Germinant Receptor, and Comparison with That of Operons Encoding Other Germinant Receptors

2006 ◽  
Vol 188 (11) ◽  
pp. 4131-4136 ◽  
Author(s):  
Takao Igarashi ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Type Strain ◽  
Wild Type Strain ◽  
Dipicolinic Acid ◽  
Mrna Levels ◽  
Wild Type ◽  
Putative Promoter ◽  
Rapid Fall ◽  
Sequence Similarities

ABSTRACT The gerA, gerB, and gerK operons, which encode germinant receptors in spores of Bacillus subtilis, were transcribed only in sporulation, and their mRNA levels peaked initially ∼3 h before the initiation of accumulation of the spore's dipicolinic acid. After a rapid fall, levels of these mRNAs peaked again ∼5 h later. In one wild-type strain (PS832), gerA mRNA was the most abundant, with levels of gerB and gerK mRNAs ∼50% of that of gerA mRNA, whereas gerB mRNA was the most abundant in another wild-type strain (PY79). The synthesis of gerK mRNA in sporulation was abolished by loss of the forespore-specific RNA polymerase sigma factor, σG, and induction of σG synthesis in vegetative cells led to synthesis of gerK mRNA. SpoVT, a regulator of σG-dependent gene expression, repressed gerK expression. The gerK promoter showed sequence similarities to σG-dependent promoters, and deletion of elements of this putative promoter abolished gerK expression in sporulation.

scholarly journals Effects of Major Spore-Specific DNA Binding Proteins on Bacillus subtilis Sporulation and Spore Properties

2000 ◽  
Vol 182 (24) ◽  
pp. 6906-6912 ◽  
Author(s):  
Barbara Setlow ◽  
Kelly A. McGinnis ◽  
Katerina Ragkousi ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Mother Cell ◽  
Type Strain ◽  
Wild Type Strain ◽  
Dipicolinic Acid ◽  
Dna Binding Proteins ◽  
Specific Gene ◽  
Wild Type ◽  
Spore Proteins ◽  
Spore Outgrowth

ABSTRACT Sporulation of a Bacillus subtilis strain (termed α− β−) lacking the majority of the α/β-type small, acid-soluble spore proteins (SASP) that are synthesized in the developing forespore and saturate spore DNA exhibited a number of differences from that of the wild-type strain, including delayed forespore accumulation of dipicolinic acid, overexpression of forespore-specific genes, and delayed expression of at least one mother cell-specific gene turned on late in sporulation, although genes turned on earlier in the mother cell were expressed normally in α− β− strains. The sporulation defects in α− β− strains were corrected by synthesis of chromosome-saturating levels of either of two wild-type, α/β-type SASP but not by a mutant SASP that binds DNA poorly. Spores from α− β− strains also exhibited less glutaraldehyde resistance and slower outgrowth than did wild-type spores, but at least some of these defects in α− β− spores were abolished by the synthesis of normal levels of α/β-type SASP. These results indicate that α/β-type SASP may well have global effects on gene expression during sporulation and spore outgrowth.

NEW LOCI IN DICTYOSTELIUM DISCOIDEUM DETERMINING PIGMENT FORMATION AND GROWTH ON BACILLUS SUBTILIS

Genetics ◽  
1980 ◽  
Vol 96 (1) ◽  
pp. 115-123
Author(s):  
James H Morrissey ◽  
Steven Wheeler ◽  
William F Loomis
Keyword(s):  
Bacillus Subtilis ◽  
Dictyostelium Discoideum ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Linkage Groups ◽  
New Mutation ◽  
Pigment Formation ◽  
Complementation Groups ◽  
First Time

ABSTRACT Seventeen independently isolated pigmentless (white) mutations in Dictyostelium discoideum are all recessive and fall into three complementation groups identifying two new whi loci in addition to the previously characterized whiA locus. whiB and whiC map to linkage groups III and IV, respectively. In addition, it was discovered that our laboratory stock of NC4, the wild-type strain from which these mutants were derived, has spontaneously lost the ability to grow on Bacillus subtilis. This new mutation, bsgB500, maps to linkage group VII and is not allelic to bsgA. bsgB500 is the first spontaneously derived mutation in D. discoideum that can be used to select heterozygous diploids, and for the first time allows genetic analysis to be routinely performed on strains derived from an unmutagenized background.

scholarly journals Selective Pressure for Biofilm Formation in Bacillus subtilis: Differential Effect of Mutations in the Master Regulator SinR on Bistability

mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Jan Kampf ◽  
Jan Gerwig ◽  
Kerstin Kruse ◽  
Robert Cleverley ◽  
Miriam Dormeyer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Biofilm Formation ◽  
Type Strain ◽  
Wild Type Strain ◽  
Selective Pressure ◽  
Wild Type ◽  
Master Regulator ◽  
Form Complex ◽  
Content Type

ABSTRACT Biofilm formation by Bacillus subtilis requires the expression of genes encoding enzymes for extracellular polysaccharide synthesis and for an amyloid-like protein. The master regulator SinR represses all the corresponding genes, and repression of these key biofilm genes is lifted when SinR interacts with its cognate antagonist proteins. The YmdB phosphodiesterase is a recently discovered factor that is involved in the control of SinR activity: cells lacking YmdB exhibit hyperactive SinR and are unable to relieve the repression of the biofilm genes. In this study, we have examined the dynamics of gene expression patterns in wild-type and ymdB mutant cells by microfluidic analysis coupled to time-lapse microscopy. Our results confirm the bistable expression pattern for motility and biofilm genes in the wild-type strain and the loss of biofilm gene expression in the mutant. Moreover, we demonstrated dynamic behavior in subpopulations of the wild-type strain that is characterized by switches in sets of the expressed genes. In order to gain further insights into the role of YmdB, we isolated a set of spontaneous suppressor mutants derived from ymdB mutants that had regained the ability to form complex colonies and biofilms. Interestingly, all of the mutations affected SinR. In some mutants, large genomic regions encompassing sinR were deleted, whereas others had alleles encoding SinR variants. Functional and biochemical studies with these SinR variants revealed how these proteins allowed biofilm gene expression in the ymdB mutant strains. IMPORTANCE Many bacteria are able to choose between two mutually exclusive lifestyles: biofilm formation and motility. In the model bacterium Bacillus subtilis, this choice is made by each individual cell rather than at the population level. The transcriptional repressor SinR is the master regulator in this decision-making process. The regulation of SinR activity involves complex control of its own expression and of its interaction with antagonist proteins. We show that the YmdB phosphodiesterase is required to allow the expression of SinR-repressed genes in a subpopulation of cells and that such subpopulations can switch between different SinR activity states. Suppressor analyses revealed that ymdB mutants readily acquire mutations affecting SinR, thus restoring biofilm formation. These findings suggest that B. subtilis cells experience selective pressure to form the extracellular matrix that is characteristic of biofilms and that YmdB is required for the homeostasis of SinR and/or its antagonists.

scholarly journals Characterization of a Wild-Type Strain ofFrancisella TularensisIsolated from a Cat

2004 ◽  
Vol 16 (5) ◽  
pp. 374-381 ◽  
Author(s):  
Thomas J. Inzana ◽  
Gretchen E. Glindemann ◽  
Gerald Snider ◽  
Susan Gardner ◽  
Lisa Crofton ◽  
...  
Keyword(s):  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type

scholarly journals A Mutation in the 3-Phosphoglycerate Kinase Gene Allows Anaerobic Growth of Bacillus subtilis in the Absence of ResE Kinase

1999 ◽  
Vol 181 (22) ◽  
pp. 7087-7097 ◽  
Author(s):  
Michiko M. Nakano ◽  
Yi Zhu ◽  
Koki Haga ◽  
Hirofumi Yoshikawa ◽  
Abraham L. Sonenshein ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Type Strain ◽  
Wild Type Strain ◽  
Response Regulator ◽  
Anaerobic Respiration ◽  
Phosphoglycerate Kinase ◽  
Anaerobic Growth ◽  
Wild Type ◽  
Kinase Gene ◽  
Glycolytic Intermediate

ABSTRACT The Bacillus subtilis ResD-ResE two-component signal transduction system is essential for aerobic and anaerobic respiration. A spontaneous suppressor mutant that expresses ResD-controlled genes and grows anaerobically in the absence of the ResE histidine kinase was isolated. In addition, aerobic expression of ResD-controlled genes in the suppressed strain was constitutive and occurred at a much higher level than that observed in the wild-type strain. The suppressing mutation, which mapped to pgk, the gene encoding 3-phosphoglycerate kinase, failed to suppress a resDmutation, suggesting that the suppressing mutation creates a pathway for phosphorylation of the response regulator, ResD, which is independent of the cognate sensor kinase, ResE. The pgk-1mutant exhibited very low but measurable 3-phosphoglycerate kinase activity compared to the wild-type strain. The results suggest that accumulation of a glycolytic intermediate, probably 1,3-diphosphoglycerate, is responsible for the observed effect of thepgk-1 mutation on anaerobiosis of resE mutant cells.

scholarly journals Characterization of melanin produced by a wild-type strain of Bacillus thuringiensis

2004 ◽  
Vol 50 (4) ◽  
pp. 183-188 ◽  
Author(s):  
Yuehua Chen ◽  
Yinyue Deng ◽  
Jinhong Wang ◽  
Jun Cai ◽  
Gaixin Ren
Keyword(s):  
Bacillus Thuringiensis ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type

Purification and characterization of a β-glucosidase from Streptomyces lividans 66

1990 ◽  
Vol 36 (1) ◽  
pp. 53-56 ◽  
Author(s):  
Anca Mihoc ◽  
Dieter Kluepfel
Keyword(s):  
High Performance Liquid Chromatography ◽  
Molecular Sieve ◽  
Type Strain ◽  
High Performance ◽  
Wild Type Strain ◽  
Streptomyces Lividans ◽  
Wild Type ◽  
Purification And Characterization ◽  
Anion Exchange Column

An intracellular β-1, 4-D-glucosidase (EC 3.2.1.21) was isolated from the mutant strain HP-3 of Streptomyces lividans 66 which produced about 12 times more enzyme than the wild-type strain. The purification was carried out by anion exchange column chromatography followed by high-performance liquid chromatography on DEAE and on molecular sieve columns. The enzyme is glycosylated and has an apparent Mr of 51 000 and a pI of 4.3. Its activity was optimal at pH 6.5 and at a temperature of 40 °C. The Km and the Vmax on cellobiose were 3.1 mM and 65.6 μmol min−1 mg−1 of enzyme. Key words: β-glucosidase, Streptomyces lividans, purification, characterization.

scholarly journals Bacillus subtilis Phosphorylated PhoP: Direct Activation of the EσA- and Repression of the EσE-Responsive phoB-PS+V Promoters during Pho Response

2005 ◽  
Vol 187 (15) ◽  
pp. 5166-5178 ◽  
Author(s):  
Wael R. Abdel-Fattah ◽  
Yinghua Chen ◽  
Amr Eldakak ◽  
F. Marion Hulett
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Subtilis ◽  
Type Strain ◽  
Wild Type Strain ◽  
Pho Regulon ◽  
Dependent Manner ◽  
Wild Type ◽  
Phosphate Deprivation

ABSTRACT The phoB gene of Bacillus subtilis encodes an alkaline phosphatase (PhoB, formerly alkaline phosphatase III) that is expressed from separate promoters during phosphate deprivation in a PhoP-PhoR-dependent manner and at stage two of sporulation under phosphate-sufficient conditions independent of PhoP-PhoR. Isogenic strains containing either the complete phoB promoter or individual phoB promoter fusions were used to assess expression from each promoter under both induction conditions. The phoB promoter responsible for expression during sporulation, phoB-PS, was expressed in a wild-type strain during phosphate deprivation, but induction occurred >3 h later than induction of Pho regulon genes and the levels were approximately 50-fold lower than that observed for the PhoPR-dependent promoter, phoB-PV. EσE was necessary and sufficient for PS expression in vitro. PS expression in a phoPR mutant strain was delayed 2 to 3 h compared to the expression in a wild-type strain, suggesting that expression or activation of σE is delayed in a phoPR mutant under phosphate-deficient conditions, an observation consistent with a role for PhoPR in spore development under these conditions. Phosphorylated PhoP (PhoP∼P) repressed PS in vitro via direct binding to the promoter, the first example of an EσE-responsive promoter that is repressed by PhoP∼P. Whereas either PhoP or PhoP∼P in the presence of EσA was sufficient to stimulate transcription from the phoB-PV promoter in vitro, roughly 10- and 17-fold-higher concentrations of PhoP than of PhoP∼P were required for PV promoter activation and maximal promoter activity, respectively. The promoter for a second gene in the Pho regulon, ykoL, was also activated by elevated concentrations of unphosphorylated PhoP in vitro. However, because no Pho regulon gene expression was observed in vivo during Pi -replete growth and PhoP concentrations increased only threefold in vivo during phoPR autoinduction, a role for unphosphorylated PhoP in Pho regulon activation in vivo is not likely.

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