Relationship between the tricarboxylic acid cycle enzymes and sporulation of B. subtilis

Hanson, Richard S.; Blicharska, J.; Szulmajster, Jekisiel

doi:10.1016/0006-291x(64)90290-6

Cited by 104 publications

(46 citation statements)

References 4 publications

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“…We assume that the ammonium sulfate treatment removes a component from CaLP, perhaps another protein, which blocks the antigenic as well as the phosphodiesterasebinding process. This apparent activation by ammonium (16) were harvested at t0,I tl1I t3, and t5 (the subscript denotes hours after the end of exponential growth). Extracts were purified through step 5 (Table 1), quantitatively applied to phenothiazine columns, and eluted with EGTA.…”

Section: Resultsmentioning

confidence: 99%

Purification and properties of an intracellular calmodulinlike protein from Bacillus subtilis cells

1991

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Although calcium ions are crucial in a variety of bacterial processes, including spore development, reports of calmodulin in procaryotes have been few. We have purified to homogeneity a calmodulinlike protein (CaLP) from sporulating cells of Bacillus subtilis grown in a chemically defined sporulation medium; purification involved heat treatment, fractionation with ammonium sulfate, affinity chromatography, and gel filtration on high-performance columns. The protein was eluted from a phenothiazine affinity column in a calcium iondependent manner, stained poorly with Coomassie blue and silver stain dyes, bound poorly to nitrocellulose filters, and was not an inhibitor of the major intracellular serine proteinase. It stimulated bovine brain phosphodiesterase in a dose-and Ca2+-dependent manner and stimulated NAD kinase from peas in a dosedependent manner. The B. subtilis calmodulin reacted with anti-bovine brain calmodulin antibodies in enzymelinked immunoabsorbance assays. The amino acid composition data showed it to be distinctly different from eucaryotic calmodulins, having particularly high levels of serine and glycine. The pl of the protein was estimated to be 4.9 to 5.0. The molecular weight was estimated to be 23,000 or 25,000, based on amino acid composition and detergent gel electrophoresis, respectively. The protein reacted with rhodamine isothiocyanate, which blocked its enzyme-activating capacity and greatly increased its electrophoretic mobility and Coomassie dye-binding ability.

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Section: Resultsmentioning

confidence: 99%

Purification and properties of an intracellular calmodulinlike protein from Bacillus subtilis cells

1991

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“…Bacteria were grown in the following media: B. brevis ATCC in the medium of Hanson et al (7); B. polymyxa ATCC 25901 in the medium of Stansly, Shepherd, and White (8); and B. subtilis ATCC 6051 in LB broth (1% Bacto tryptone, 0.5% yeast extract, and 0.5% NaCl at pH 7.0). Growth was at 370 with vigorous aeration in a New Brunswick Microferm or Fermacell fermentor, and the cells were harvested during midexponential growth with a refrigerated Sharples Supercentrifuge.…”

Section: Methodsmentioning

confidence: 99%

Nucleotide-Dependent Inactivation of RNA Polymerase from Bacillus brevis

Sarkar¹,

Paulus²

1972

Proc. Natl. Acad. Sci. U.S.A.

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RNA polymerase has been purified from vegetative cells of Bacillus brevis and resolved into 'core' enzyme and sigma factor. The purified enzyme is rapidly inactivated by incubation at low temperatures in the presence of 1-2 mM ATP, dATP, or NAD+, while other nucleotides at this concentration have little or no effect. Inactivation is not accompanied by the incorporation of an adenylyl or phosphoryl moiety into RNA polymerase; nevertheless, it is essentially irreversible. DNA, high concentrations of glycerol, as well as low concentrations (1 mM) of orthophosphate protect RNA polymerase from the nucleotide-dependent inactivation.A similar inactivation of RNA polymerase in the presence of ATP is observed with crude preparations from Bacillus subtilis and Bacillus polymyxa. This phenomenon may represent a novel mode of regulation of transcription that does not involve a covalent modification of RNA polymerase or its interaction with other protein factors, but rather is due to a structural transition to an inactive form induced by small molecules.The transcription of specific genes by bacterial RNA polymerases is known to be modulated by protein factors that interact either with the enzyme itself or with the DNA template (1-3). Under conditions of drastic physiological change, as upon bacteriophage infection or during sporulation, a more general control of transcription is seen that often involves the covalent modification of RNA polymerase (4, 5). Between these two extreme types of regulation lies the noncovalent modification of RNA polymerase by small molecules, such as the inhibition by peptide antibiotics during the early stages of sporulation (6). In this paper, we describe a phenomenon that may also belong in this category-although its physiological significance is still obscure-namely, the inactivation of RNA polymerase from Bacillus brevis in the presence of specific nucleotides. precipitated with ammonium sulfate (70% saturation), and redissolved in Buffer A containing 50% glycerol for storage at -20°(Fraction IV). The purification procedure is summarized in Table 1. MATERIALS AND METHODS Growth ofCrude preparations of RNA polymerase were obtained from sonic extracts of B. polymyxa and B. brevis by centrifugation at 100,000 X g for 1 hr and fractionation with ammonium sulfate.Enzyme Assays. RNA polymerase was assayed at 370 for 15 min in the presence of 0.1 mM [5-3H]UTP (10 Ci/mol), 1 mM (each) of ATP, GTP, and CTP, 10 mM MgCl2, 2 mM MnCl2, 10 mM 2-mercaptoethanol, bovine serum albumin (0.5 mg/ml), and DNA (0.1 mg/ml of salmon-sperm DNA unless otherwise indicated), in a final volume of 0.2 ml. The reaction was terminated by the addition of cold trichloroacetic acid (0.3 N), and the precipitate was washed with trichloroacetic acid, then with ethanol, on discs of glass-fiber filter paper (Whatman GF/C). The radioactivity was determined in a liquid scintillation spectrometer in a toluene-based scintillation fluid. 1 Unit of RNA polymerase activity is defined as the amount of enzyme that catalyzes the inc...

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“…Kreisel (1972) showed that a few days after all ants are removed from the fungus garden, the pH gradually increases and the garden is quickly overgrown by opportunistic fungi and bacteria. Acetic acid and fatty acids are general metabolites of many micro-organisms and most Bacillus species, including Bacillus subtilis, can break down acetic acid to CO 2 and H 2 O (Hanson et al, 1963;1964). The relatively strong sensitivity of the bacteria to this compound is therefore somewhat surprising.…”

Section: Function Of the Specific Classes Of Metapleural Gland Compoundsmentioning

confidence: 99%

Variable sensitivity of fungi and bacteria to compounds produced by the metapleural glands of leaf-cutting ants

Bot

Ortius-Lechner

Finster³

et al. 2002

Insectes soc.

129

131

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Relationship between the tricarboxylic acid cycle enzymes and sporulation of B. subtilis

Cited by 104 publications

References 4 publications

Purification and properties of an intracellular calmodulinlike protein from Bacillus subtilis cells

Purification and properties of an intracellular calmodulinlike protein from Bacillus subtilis cells

Nucleotide-Dependent Inactivation of RNA Polymerase from Bacillus brevis

Variable sensitivity of fungi and bacteria to compounds produced by the metapleural glands of leaf-cutting ants

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