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

B. K. Rima; I. Takahashi

doi:10.1139/o73-161

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

Canadian Journal of Biochemistry ◽

10.1139/o73-161 ◽

1973 ◽

Vol 51 (9) ◽

pp. 1219-1224 ◽

Cited By ~ 7

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.

Download Full-text

BACTERIOPHAGE T4 ENDONUCLEASES II AND IV, OPPOSITELY AFFECTED BY dCMP HYDROXYMETHYLASE ACTIVITY, HAVE DIFFERENT ROLES IN THE DEGRADATION AND IN THE RNA POLYMERASE-DEPENDENT REPLICATION OF T4 CYTOSINE-CONTAINING DNA

Genetics ◽

10.1093/genetics/114.3.669 ◽

1986 ◽

Vol 114 (3) ◽

pp. 669-685

Author(s):

Karin Carlson ◽

Aud Ȗvervatin

Keyword(s):

Rna Polymerase ◽

Dna Synthesis ◽

Bacteriophage T4 ◽

The Other ◽

Wild Type ◽

E Coli ◽

Phage Dna ◽

Dna Template ◽

Do So

ABSTRACT Bacteriophage T4 mutants defective in gene 56 (dCTPase) synthesize DNA where cytosine (Cyt) partially or completely replaces hydroxymethylcytosine (HmCyt). This Cyt-DNA is degraded in vivo by T4 endonucleases II and IV, and by the exonuclease coded or controlled by genes 46 and 47.—Our results demonstrate that T4 endonuclease II is the principal enzyme initiating degradation of T4 Cyt-DNA. The activity of endonuclease IV, but not that of endonuclease II, was stimulated in the presence of a wild-type dCMP hydroxymethylase, also when no HmCyt was incorporated into phage DNA, suggesting the possibility of direct endonuclease IV-dCMP hydroxymethylase interactions. Endonuclease II activity, on the other hand, was almost completely inhibited in the presence of very small amounts of HmCyt (3-9% of total Cyt + HmCyt) in the DNA. Possible mechanisms for this inhibition are discussed.—The E. coli RNA polymerase modified by the products of T4 genes 33 and 55 was capable of initiating DNA synthesis on a Cyt-DNA template, although it probably cannot do so on an HmCyt template. In the presence of an active endonuclease IV, Cyt-DNA synthesis was arrested 10-30 min after infection, probably due to damage to the template. Cyt-DNA synthesis dependent on the unmodified (33 - 55 -) RNA polymerase was less sensitive to endonuclease IV action.

Download Full-text

Genetic and physiological studies of bacteriophage T5 2. The relationship between phage DNA synthesis and protein synthesis in T5-infected cells

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(71)90677-2 ◽

1971 ◽

Vol 43 (4) ◽

pp. 735-740 ◽

Cited By ~ 8

Author(s):

Herbert E. Hendrickson ◽

D. James McCorquodale

Keyword(s):

Protein Synthesis ◽

Dna Synthesis ◽

Phage Dna ◽

Infected Cells ◽

Physiological Studies ◽

Bacteriophage T5 ◽

The Relationship

Download Full-text

THE EFFECT OF ULTRAVIOLET LIGHT ON THE PRODUCTION OF BACTERIAL VIRUS PROTEIN

The Journal of General Physiology ◽

10.1085/jgp.40.4.521 ◽

1957 ◽

Vol 40 (4) ◽

pp. 521-531 ◽

Cited By ~ 31

Author(s):

Itaru Watanabe

Keyword(s):

Infected Cell ◽

Dna Synthesis ◽

Ultraviolet Light ◽

Specific Protein ◽

Virus Protein ◽

Antigenic Protein ◽

Phage T4 ◽

Phage Dna ◽

Phage Protein ◽

Infected Cells

The amount of phage-specific protein in T2-infected bacteria growing in a medium containing radiosulfur, S35, has been studied by measuring the radioactivity in specific antiphage serum precipitates of lysates. In the course of normal infection, non-infective phage antigen has been found to make its first intracellular appearance shortly before the end of the eclipse period, in agreement with the findings of Maaløe and Symonds with phage T4. No such phage antigen is produced either in bacteria infected with UV-inactivated T2 or in T2-infected bacteria whose survival as an infective center has been destroyed by UV irradiation during the early stages of the eclipse period. If the infected bacteria are UV-irradiated only at later stages of the eclipse period however, then phage antigenic protein continues to be synthesized in those infected cells in which DNA synthesis and, a fortiori, production of infective progeny have been almost completely suppressed. It is concluded from these results that once the mechanism for formation of phage-specific protein has been established within the infected cell under the influence of the parental DNA, synthesis of phage-specific protein can continue independently of the synthesis of phage DNA. The possibility that the phage DNA controls the specificity of the phage protein indirectly through substances other than DNA is discussed.

Download Full-text

WATER FLOW THROUGH FROG GASTRIC MUCOSA

The Journal of General Physiology ◽

10.1085/jgp.39.4.535 ◽

1956 ◽

Vol 39 (4) ◽

pp. 535-551 ◽

Cited By ~ 76

Author(s):

Richard P. Durbin ◽

Heddy Frank ◽

A. K. Solomon

Keyword(s):

Protein Synthesis ◽

Dna Synthesis ◽

Time Interval ◽

Appreciable Effect ◽

E Coli ◽

Evolution Of Resistance ◽

Infected Cells ◽

Flow Through ◽

Phage Growth

To elucidate the role of protein synthesis in DNA formation, E. coli R2 infected with phage T2 was studed as a model, employing chloramphenicol to inhibit protein synthesis. The following results were obtained. 1. Chloramphenicol inhibited protein synthesis but not synthesis of nucleic acids in uninfected bacteria. 2. Studies of the effect of chloramphenicol on phage maturation indicated a delay of 2 minutes between time of addition and cessation of phage growth. 3. The increase of DNA in phage-infected bacteria was completely suppressed by the addition of chloramphenicol within 2 minutes following infection. Addition at later times showed progressively less inhibitory action depending upon the time interval, and addition after the 10th or 12th minute showed no appreciable effect on DNA synthesis despite the cessation of intracellular phage formation and protein synthesis. 4. When chloramphenicol was added to infected cells the increase of resistance to UV stopped within 2 minutes, whether or not DNA synthesis continued. Thus evolution of resistance paralleled the rate of DNA synthesis achieved, but not the amount of DNA accumulated. 5. We conclude that in infected bacteria, protein synthesis is necessary to initiate DNA synthesis but is not essential for its continuation. The resistance to UV that characterizes infected cells near the midpoint of the latent period is not due to accumulation of DNA, but depends on some chloramphenicol-sensitive process (probably protein synthesis) completed at about the time the rate of DNA synthesis becomes maximal.

Download Full-text

Adenylation and ADP-ribosylation in the mouse 1-cell embryo

Development ◽

10.1242/dev.49.1.139 ◽

1979 ◽

Vol 49 (1) ◽

pp. 139-152

Author(s):

R. J. Young ◽

K. Sweeney

Keyword(s):

Dna Synthesis ◽

Snake Venom ◽

Trichloroacetic Acid ◽

Actinomycin D ◽

Adp Ribosylation ◽

Snake Venom Phosphodiesterase ◽

Insoluble Material ◽

Cell Embryo

The incorporation of [3H]adenosine into cold trichloroacetic acid (TCA) insoluble material by the mouse 1-cell embryo has been studied. Incorporation of label was high immediately after fertilization, then decreased over the next 7 h with the sharpest decline occurring 3—5 h after fertilization. A small maximum was observed at the time of pronuclear DNA synthesis. Actinomycin D at a concentration which inhibited the cleavage of 1-cell embryos by 50 % had little effect on this incorporation, which in the period 1—6 h post-fertilization was shown by autoradiography to be confined to the ooplasm of the newly fertilized ovum. [3H]Adenosine and poly ([H]A) were released from embryo RNA labelled 1—3 h after fertilization with [3H]adenosine by digestion with a mixture of ribonucleases A and T1. The poly ([3H]A) segments were hydrolysed by alkali to 3′-[3H]AMP and [3H]adenosine ([3H]AMP/[3H]adenosine = 5/1), and by snake venom phosphodiesterase to 5′-[3H]AMP but very little [3H]adenosine. These results suggest that adenylation of RNA occurs soon after fertilization, that this is a cytoplasmic event, and that most of the newly synthesized poly ([3H]A) segments are joined to pre-existing poly (A) tracts. The unusual polynucleotide, poly (ADP-ribose), identified by its resistance to alkali and the release of 2′-(5″-phosphoribosyl)-5′[3H]AMP on incubation with snake venom phosphodiesterase, was also found in the ribonuclease digest.

Download Full-text

THE EFFECT OF CHLORAMPHENICOL ON DEOXYRIBONUCLEIC ACID SYNTHESIS AND THE DEVELOPMENT OF RESISTANCE TO ULTRAVIOLET IRRADIATION IN E. COLI INFECTED WITH BACTERIOPHAGE T2

The Journal of General Physiology ◽

10.1085/jgp.39.4.553 ◽

1956 ◽

Vol 39 (4) ◽

pp. 553-565 ◽

Cited By ~ 90

Author(s):

Jun-ichi Tomizawa ◽

Sumiko Sunakawa

Keyword(s):

Protein Synthesis ◽

Dna Synthesis ◽

Acid Synthesis ◽

Time Interval ◽

Appreciable Effect ◽

E Coli ◽

Development Of Resistance ◽

Deoxyribonucleic Acid Synthesis ◽

Infected Cells ◽

Phage Growth

To elucidate the role of protein synthesis in DNA formation, E. coli R2 infected with phage T2 was studied as a model, employing chloramphenicol to inhibit protein synthesis. The following results were obtained. 1. Chloramphenicol inhibited protein synthesis but not synthesis of nucleic acids in uninfected bacteria. 2. Studies of the effect of chloramphenicol on phage maturation indicated a delay of 2 minutes between time of addition and cessation of phage growth. 3. The increase of DNA in phage-infected bacteria was completely suppressed by the addition of chloramphenicol within 2 minutes following infection. Addition at later times showed progressively less inhibitory action depending upon the time interval, and addition after the 10th or 12th minute showed no appreciable effect on DNA synthesis despite the cessation of intracellular phage formation and protein synthesis. 4. When chloramphenicol was added to infected cells the increase of resistance to UV stopped within 2 minutes, whether or not DNA synthesis continued. Thus evolution of resistance paralleled the rate of DNA synthesis achieved, but not the amount of DNA accumulated. 5. We conclude that in infected bacteria, protein synthesis is necessary to initiate DNA synthesis but is not essential for its continuation. The resistance to UV that characterizes infected cells near the midpoint of the latent period is not due to accumulation of DNA, but depends on some chloramphenicol-sensitive process (probably protein synthesis) completed at about the time the rate of DNA synthesis becomes maximal.

Download Full-text

Use of Antibody to aid Visualization of Protein at the Termini of Bacteriophage Ø29 DNA

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600002385 ◽

1980 ◽

Vol 38 ◽

pp. 540-541

Author(s):

Dwight Anderson ◽

Charlene Peterson ◽

Gursaran Notani ◽

Bernard Reilly

Keyword(s):

Electron Microscopy ◽

Bacillus Subtilis ◽

Dna Synthesis ◽

Restriction Endonuclease ◽

Protein Complex ◽

Protein Product ◽

Viral Dna ◽

Small Proteins ◽

Antibody Labeling ◽

Subtilis Bacteriophage

The protein product of cistron 3 of Bacillus subtilis bacteriophage Ø29 is essential for viral DNA synthesis and is covalently bound to the 5’-termini of the Ø29 DNA. When the DNA-protein complex is cleaved with a restriction endonuclease, the protein is bound to the two terminal fragments. The 28,000 dalton protein can be visualized by electron microscopy as a small dot and often is seen only when two ends are in apposition as in multimers or in glutaraldehyde-fixed aggregates. We sought to improve the visibility of these small proteins by use of antibody labeling.

Download Full-text

EFFECT OF ACTINOMYCIN D ON TSH-INDUCED STIMULATION OF THYROIDAL PROTEIN SYNTHESIS IN THE RAT

Acta Endocrinologica ◽

10.1530/acta.0.0770064 ◽

1974 ◽

Vol 77 (1) ◽

pp. 64-70 ◽

Cited By ~ 9

Author(s):

Gustav Wägar

Keyword(s):

Protein Synthesis ◽

Actinomycin D ◽

The Other ◽

Leucine Incorporation ◽

Short Term ◽

Other Hand ◽

Thyroid Glands ◽

Translational Level ◽

Stimulation Of

ABSTRACT Whether the short-term regulation of thyroidal protein synthesis by TSH occurs at the transcriptional or the translational level was tested by measuring the effect of actinomycin D (act D) on the TSH-induced stimulation of L-14C-leucine incorporation into the thyroidal proteins of rats. TSH was injected 6 h before the rats were killed. The thyroid glands were then removed and incubated in vitro in the presence of L-14C-leucine for 2 h. The pronounced stimulation of leucine incorporation in the TSH-treated animals was depressed as compared with controls but still significant even when the animals had been pre-treated with 100 μg act D 24 and 7 h before sacrifice. On the other hand, act D strongly decreased incorporation of 3H-uridine into RNA. Short-term regulation of thyroidal protein synthesis by TSH appears to be partly but not wholly dependent on neosynthesis of RNA. Hence regulation may partly occur at the translation level of protein synthesis.

Download Full-text