Effects of actinomycin D and 5-fluorouracil on the formation of enzymes in Bacillus subtilis

1965 ◽  
Vol 103 (2) ◽  
pp. 311-318 ◽  
Author(s):  
Kiyoshi Kadowaki ◽  
Junko Hosoda ◽  
Bunji Maruo
Keyword(s):  
Bacillus Subtilis ◽  
Actinomycin D

Synthesis of acid-soluble spore proteins by Bacillus subtilis

1982 ◽  
Vol 152 (3) ◽  
pp. 1117-1125
Author(s):  
J M Leventhal ◽  
G H Chambliss
Keyword(s):  
Bacillus Subtilis ◽  
Maximum Rate ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Sodium Dodecyl ◽  
Molecular Weights ◽  
Spore Proteins ◽  
Major Acid

The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phosphatase synthesis began at about t2, and refractile spores were first observed between t7 and t8. In vivo- and in vitro-synthesized ASSPs comigrated in sodium dodecyl sulfate-polyacrylamide gels. Their molecular weights were 4,600 (alpha and beta) and 11,000 (gamma). The average half-life of the ASSP messages was 11 min when either rifampin (10 micrograms/ml) or actinomycin D (1 microgram/ml) was used to inhibit RNA synthesis.

scholarly journals Commitment to sporulation in Bacillus subtilis and its relationship to development of actinomycin resistance

1969 ◽  
Vol 113 (1) ◽  
pp. 29-37 ◽  
Author(s):  
Janet M. Sterlini ◽  
J. Mandelstam
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Messenger Rna ◽  
Actinomycin D ◽  
Dipicolinic Acid ◽  
Vegetative State ◽  
Single Point

1. Experiments to determine the point of commitment to sporulation were carried out by restoring nutrients at different times to suspensions of sporulating Bacillus subtilis. 2. No single point of commitment to the process as a whole was found. Instead, the cells became committed in turn to the following successive events connected with sporulation: formation of alkaline phosphatase, development of refractility, synthesis of dipicolinic acid and development of heat-resistance. 3. Each point of commitment was followed within about 30min. by a period in which the event concerned ceased to be inhibited by actinomycin D. 4. The implication of these results is that each point of commitment is probably due to the formation of a species of long-lived messenger RNA and that, in any case, sporulation is regulated at the level of both transcription and translation. 5. It is also shown that sporulation and growth are perhaps not mutually exclusive functions and that histidase, an enzyme typical of the vegetative state, can be induced in sporulating suspensions.

The Synthesis of Nucleic Acids in Bacillus subtilis Infected with Phage PBS 1

1973 ◽  
Vol 51 (9) ◽  
pp. 1219-1224 ◽  
Author(s):  
B. K. Rima ◽  
I. Takahashi
Keyword(s):  
Bacillus Subtilis ◽  
Nucleic Acids ◽  
Dna Synthesis ◽  
Trichloroacetic Acid ◽  
Actinomycin D ◽  
The Other ◽  
Phage Dna ◽  
Infected Cells ◽  
Phage Growth ◽  
A New Species

Unlike other phage systems, the development of PBS 1 was found to be insensitive to rifamycin SV. The incorporation of 3H-uridine into trichloroacetic acid precipitable and alkali-labile material (RNA), in PBS-1-infected cells, was greatly reduced by rifamycin. Observations that RNA synthesized in the presence of rifamycin was hybridizable exclusively with the phage DNA and that actinomycin D inhibited the phage growth indicated that the synthesis of a new species of RNA was required for the development of PBS 1. The host DNA synthesis was reduced to a very low level 5 min after infection. The phage DNA synthesis was also apparently reduced markedly by rifamycin when determined with 3H-uridine as labelling material. On the other hand, rifamycin did not affect the incorporation of 3H-deoxycytidine into the phage DNA, suggesting that phage DNA synthesis was in fact insensitive to rifamycin. It is not clear how rifamycin inhibits the incorporation of 3H-uridine into nucleic acids in PBS-1-infected cells.

ACCUMULATION OF GUANINE NUCLEOTIDES IN THE ACID-SOLUBLE FRACTION OF BACILLUS SUBTILIS INHIBITED BY ACTINOMYCIN D

1966 ◽  
Vol 44 (1) ◽  
pp. 9-18 ◽  
Author(s):  
G. G. Jacoli ◽  
S. H. Zbarsky
Keyword(s):  
Bacillus Subtilis ◽  
Actinomycin D ◽  
Soluble Fraction ◽  
Guanosine Triphosphate ◽  
Control Material ◽  
Guanosine Diphosphate ◽  
Inosinic Acid ◽  
Cytidine Diphosphate ◽  
Layer Chromatography ◽  
Soluble Fractions

Acid-soluble extracts were made of cells of Bacillus subtilis grown on a medium containing glycine-2-14C as a labeled precursor. Paper chromatography of the extracts with propan-2-ol:H2O:NH3as solvent and radioautography of the chromatograms showed that if growth of the cells was inhibited by including actinomycin D in the medium, two radioactive ultraviolet-absorbing components appeared close to the origin of the chromatograms. Only traces were present in control cells not inhibited by the antibiotic. Spectrophotometry of eluted material indicated that these were compounds of guanine, and electrophoresis and phosphorus determinations provided evidence that they were guanosine triphosphate and guanosine diphosphate. Further evidence for this was obtained y comparing them with authentic nucleotides by thin-layer chromatography and by showing that treatment with phosphomonoesterase liberated guanosine. The presence of other components in the acid-soluble fractions was observed and two have been identified tentatively. One may be cytidine diphosphate, which is more strongly radioactive in the control cells. The second, which appears to be radioactive inosinic acid, was present in the inhibited cells but not in the control material.

scholarly journals Lethal effect of rifampicin in Bacillus subtilis as a complicating factor in the assessment of the lifetime of messenger ribonucleic acid

1973 ◽  
Vol 134 (1) ◽  
pp. 263-270 ◽  
Author(s):  
John G. Coote ◽  
David A. Wood ◽  
Joel Mandelstam
Keyword(s):  
Bacillus Subtilis ◽  
Phase Contrast ◽  
Actinomycin D ◽  
Lethal Effect ◽  
Cell Integrity ◽  
Bactericidal Action ◽  
Physical Damage ◽  
Messenger Ribonucleic Acid ◽  
Bacillus Subtilis 168 ◽  
Synchronized Cultures

The bactericidal action of rifampicin was compared with that of chloramphenicol in growing and in sporulating cultures of Bacillus subtilis 168. Chloramphenicol kills cells only very slowly, but exposure to rifampicin kills over 95% of cells in a few minutes, causing gross physical damage, which is visible in both phase-contrast and electron microscopy. This is accompanied by a fall in O2 consumption and by lysis. Experiments with synchronized cultures showed that susceptibility to the lethal effect of rifampicin is greater when the cells are dividing. The results suggest that the synthesis of some species of RNA other than mRNA may be necessary for the maintenance of cell integrity, although experiments with actinomycin D do not altogether fit this interpretation. However, we conclude that rifampicin is too toxic to use as an antibiotic for assessing the lifetime of mRNA.

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