Effects of various inhibitory agents on sporulation of Bacillus subtilis

1982 ◽  
Vol 28 (1) ◽  
pp. 80-86 ◽  
Author(s):  
I. Takahashi ◽  
L. W. MacKenzie

Electron microscopic examinations of Bacillus subtilis cells revealed that relatively high concentrations of carbon sources blocked sporulation at stage 0 in most cells. Both nalidixic acid and novobiocin blocked sporulation at stage 0. The cells treated with acridine orange showed the morphology of stage IV 5 h after the end of exponential growth, but no further progression was observed. Mutants that are able to sporulate in the presence of these agents had the characteristic morphological changes observed in uninhibited cultures.

1970 ◽  
Vol 118 (4) ◽  
pp. 667-676 ◽  
Author(s):  
W. M. Waites ◽  
D. Kay ◽  
I. W. Dawes ◽  
D. A. Wood ◽  
S. C. Warren ◽  
...  

A comparison was made of morphological changes and successive, mainly biochemical, marker events for sporulation in 14 asporogenous mutants. The morphological and biochemical sequences are linked so that arrested development in one is accompanied by corresponding effects in the other. Thus mutants that fail to produce both protease and antibiotic do not progress beyond stage 0, formation of alkaline phosphatase appears to be associated with the transition from stage II to stage III and glucose dehydrogenase with that from stage III to stage IV. Stage II mutants may produce `pygmy' cells or other bizarre cell-division forms. The biochemical sequence is dependent in the sense that if the occurrence of any one event is blocked that of all the succeeding events is also blocked. This has implications for biochemical models that have been proposed to explain the temporal sequence observed in spore development.


1984 ◽  
Vol 30 (4) ◽  
pp. 423-429 ◽  
Author(s):  
Dongxu Sun ◽  
I. Takahashi

By transformation analysis, a mutation (crsE1), which makes Bacillus subtilis cells able to sporulate in the presence of relatively high concentrations of glucose and other carbon sources, was mapped in the rpoBC operon. The effect of crsE1 mutation can be suppressed by another mutation in the same operon, rfm11, which confers resistance to rifamycin. Mutants carrying stv or std mutations, which are also located in the rpoBC operon, showed partial resistance to catabolites in sporulation. It appears therefore that a change in the structure or synthesis of RNA polymerase may alter the response of cells to the inhibitory effect of catabolites on sporulation.


1982 ◽  
Vol 28 (11) ◽  
pp. 1242-1251 ◽  
Author(s):  
Dongxu Sun ◽  
I. Takahashi

Using mutants of Bacillus subtilis that are able to sporulate in the presence of relatively high concentrations of various carbon sources, catabolite resistance mutations were mapped by PBS1 transduction and transformation. Catabolite resistance mutations were localized at six different loci on the chromosome of B. subtilis. The map positions of our mutants suggest that they are distinct from sacUh, catA, and scoC reported by other investigators. Relations between our findings and initiation of sporulation have been discussed.


1980 ◽  
Vol 26 (4) ◽  
pp. 420-426 ◽  
Author(s):  
Blaine L. Beaman ◽  
Kenneth C. Burtis ◽  
Roy H. Doi ◽  
James P. Yeggy ◽  
Donald P. Stahly

An electron microscopic analysis of Bacillus subtilis cells revealed that netropsin blocked sporulation ultrastructurally at stages 0–I. These observations are consistent with previous results which indicated that cells were not committed to sporulation in the presence of the drug. Further, the addition of netropsin up to stage III of sporulation prevented the normal sharp increase in dihydrodipicolinate synthase activity which results in dipicolinic acid accumulation at stage IV. The addition of netropsin after stage III had much less effect on the synthesis of dihydrodipicolinate synthase and on sporulation. Thus, morphological events in sporulation are blocked early whereas a sporulation-associated enzyme may be affected at later stages. These data indicate that netropsin affects the expression of sporulation-associated genes in a differential manner.


1973 ◽  
Vol 134 (1) ◽  
pp. 271-281 ◽  
Author(s):  
Stephen J. Brewer ◽  
Roger C. W. Berkeley

1. The control of exo-β-N-acetylglucosaminidase (EC 3.2.1.30) production by Bacillus subtilis B growing on a chemically defined medium was studied. 2. The enzyme was repressed during exponential growth by those carbon sources that enter the glycolytic pathway above the level of phosphoenolpyruvate. When exponential growth ceased as a result of low concentrations of the nitrogen, carbon or metal ion components of the medium, the enzyme was formed and its amount could be increased by the addition of cell-wall fragments as inducer. 3. The enzyme was de-repressed and could be induced during exponential growth on non-glycolytic compounds metabolized directly into pyruvate, acetyl-CoA or tricarboxylic acid cycle intermediates. 4. The major difference in the metabolism of the organism utilizing these two groups of compound was the existence of high activities of phosphoenolpyruvate carboxylase required for gluconeogenesis. 5. It is concluded that the de-repression of glucosaminidase occurs when the only principal change detected in the intermediary metabolism of the organism was the presence of high activities of phosphoenolpyruvate carboxylase. 6. When the organism was grown on media containing repressing compounds, the enzyme was only de-repressed on entry of the cells into the initial stages of sporulation, where phosphoenolpyruvate carboxylase activity, even in the presence of excess of glucose, increased in parallel with glucosaminidase, neutral proteinase and alkaline phosphatase activities. 7. These results suggest a strong link, at the level of the tricarboxylic acid cycle, between the control of phosphoenolpyruvate carboxylase and the control of the de-repression of glucosaminidase and sporulation.


Author(s):  
A.J. Mia ◽  
L.X. Oakford ◽  
T. Yorio

The amphibian urinary bladder has been used as a ‘model’ system for studies of the mechanism of action of antidiuretic hormone (ADH) in stimulating transepithelial water flow. The increase in water permeability is accompanied by morphological changes that include the stimulation of apical microvilli, mobilization of microtubules and microfilaments and vesicular membrane fusion events . It has been shown that alterations in the cytosolic calcium concentrations can inhibit ADH transmembrane water flow and induce alterations in the epithelial cell cytomorphology, including the cytoskeletal system . Recently, the subapical granules of the granular cell in the amphibian urinary bladder have been shown to contain high concentrations of calcium, and it was suggested that these cytoplasmic constituents may act as calcium storage sites for intracellular calcium homeostasis. The present study utilizes the calcium antagonist, verapamil, to examine the effect of calcium deprivation on the cytomorphological features of epithelial cells from amphibian urinary bladder, with particular emphasis on subapical granule and microfilament distribution.


Author(s):  
N. Kohyama ◽  
K. Fukushima ◽  
A. Fukami

Since the interlayer or adsorbed water of some clay minerals are quite easily dehydrated in dried air, in vacuum, or at moderate temperatures even in the atmosphere, the hydrated forms have not been observed by a conventional electron microscope(TEM). Recently, specific specimen chambers, “environmental cells(E.C.),” have been developed and confirmed to be effective for electron microscopic observation of wet specimen without dehydration. we observed hydrated forms of some clay minerals and their morphological changes by dehydration using a TEM equipped with an E.C..The E.C., equipped with a single hole copper-microgrid sealed by thin carbon-film, attaches to a TEM(JEM 7A) with an accelerating voltage 100KV and both gas pressure (from 760 Torr to vacuum) and relative humidity can be controlled. The samples collected from various localities in Japan were; tubular halloysite (l0Å) from Gumma Prefecture, sperical halloysite (l0Å) from Tochigi Pref., and intermediate halloysite containing both tubular and spherical types from Fukushima Pref..


1984 ◽  
Vol 32 (9) ◽  
pp. 973-981 ◽  
Author(s):  
B W Lubit

Previous immunocytochemical studies in which an antibody specific for mammalian cytoplasmic actin was used showed that a high concentration of cytoplasmic actin exists at neuromuscular junctions of rat muscle fibers such that the distribution of actin corresponded exactly to that of the acetylcholine receptors. Although clusters of acetylcholine receptors also are present in noninnervated rat and chick muscle cells grown in vitro, neither the mechanism for the formation and maintenance of these clusters nor the relationship of these clusters to the high density of acetylcholine receptors at the neuromuscular junction in vivo are known. In the present study, a relationship between beta-cytoplasmic actin and acetylcholine receptors in vitro has been demonstrated immunocytochemically using an antibody specific for the beta-form of cytoplasmic actin. Networks of cytoplasmic actin-containing filaments were found in discrete regions of the myotube membrane that also contained high concentrations of acetylcholine receptors; such high concentrations of acetylcholine receptors have been described in regions of membrane-substrate contact. Moreover, when primary rat myotubes were exposed to human myasthenic serum, gross morphological changes, accompanied by an apparent rearrangement of the cytoplasmic actin-containing cytoskeleton, were produced. Although whether the distribution of cytoplasmic actin-containing structures was influenced by the organization of acetylcholine receptor or vice versa cannot be determined from these studies, these findings suggest that in primary rat muscle cells grown in vitro, acetylcholine receptors and beta-cytoplasmic actin-containing structures may be somehow connected.


Microbiology ◽  
2014 ◽  
Vol 160 (2) ◽  
pp. 243-260 ◽  
Author(s):  
Öykü İrigül-Sönmez ◽  
Türkan E. Köroğlu ◽  
Büşra Öztürk ◽  
Ákos T. Kovács ◽  
Oscar P. Kuipers ◽  
...  

The lutR gene, encoding a product resembling a GntR-family transcriptional regulator, has previously been identified as a gene required for the production of the dipeptide antibiotic bacilysin in Bacillus subtilis. To understand the broader regulatory roles of LutR in B. subtilis, we studied the genome-wide effects of a lutR null mutation by combining transcriptional profiling studies using DNA microarrays, reverse transcription quantitative PCR, lacZ fusion analyses and gel mobility shift assays. We report that 65 transcriptional units corresponding to 23 mono-cistronic units and 42 operons show altered expression levels in lutR mutant cells, as compared with lutR + wild-type cells in early stationary phase. Among these, 11 single genes and 25 operons are likely to be under direct control of LutR. The products of these genes are involved in a variety of physiological processes associated with the onset of stationary phase in B. subtilis, including degradative enzyme production, antibiotic production and resistance, carbohydrate utilization and transport, nitrogen metabolism, phosphate uptake, fatty acid and phospholipid biosynthesis, protein synthesis and translocation, cell-wall metabolism, energy production, transfer of mobile genetic elements, induction of phage-related genes, sporulation, delay of sporulation and cannibalism, and biofilm formation. Furthermore, an electrophoretic mobility shift assay performed in the presence of both SinR and LutR revealed a close overlap between the LutR and SinR targets. Our data also revealed a significant overlap with the AbrB regulon. Together, these findings reveal that LutR is part of the global complex, interconnected regulatory systems governing adaptation of bacteria to the transition from exponential growth to stationary phase.


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