Inhibition of Dna-Dependent Rna Synthesis by Rifamycins

Umezawa, Hamao; Mizuno, Satoshi; Yamazaki, Hisaji; Nitta, Kazuo

doi:10.7164/antibiotics.21.234

Cited by 151 publications

(65 citation statements)

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“…coli RNA polymerase; (b) the results presented in Table 1 were obtained in the presence of rifampicin at a level which completely inhibits E. coli RNA polymerase activity [23].…”

Section: J Dunn W T M C~l I S T E R mentioning

confidence: 96%

Transcription in vitro of T3 DNA by Escherichia coli and T3 RNA Polymerases

Dunn

McAllister

Bautz

1972

European Journal of Biochemistry

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RNA isolated from Escherichia coli infected by bacteriophage T3 was sedimented through sucrose gradients. The size distributions of the mRNA species coding for three T3-specific enzymes, the S-adenosyl-L-methionine cleaving enzyme (adenosylmethioninase), phage RNA polymerase and lysozyme, were then investigated by using the fractionated RNA to program a cell-free protein-synthesizing system followed by enzymatic analysis of the translation products. Identical analyses of transcripts produced in vitro by E. coli and T3 RNA polymerases were performed and it was found that E. coli RNA polymerase, in conjuncton with termination factor (Q). was capable of synthesizing early mRNA species (mRNA for adenosylmethioninase and phage polymerase) apparently identical in size and coding capacity to those isolated from infected cells. Transcripts synthesized by the T3 RNA polymerase, while consisting predominantly of late RNA species, were found to contain functional mRNAs for adenosylmethioninase, phage RNA polymerase and lysozyme.The transcription patterns obtained are consistent with the assignment of one promotor for the region situated near the left terminus of the genome.Transcription of the T3 and T7 genomes is a t least a two-step process [I-31. Initially upon entry into the cell the left-hand segment of the phage genome is transcribed by the host transcription apparatus. This region, termed the early region, comprises approximately 20°/, of the total coding capacity of the phage genome [3] and contains a gene (gene 1) which codes for a phage-specific RNA polymerase whose expression exerts a pleiotropic positive control upon the transcription of the remaining SOo/, of the genome [4,5].I n a previous communication [6] we presented evidence that the purified phage polymerase was capable of transcribing both the early and late regions of the genome resulting in the production of RNA species which were similar in size to those observed in vivo late in the infection cycle. The production of discrete size classes of RNA by this Enzymes. RNA polymerase or RNA nucleotidyltransferase (EC 2.7.7.6); lysozyme (EC 3.2.1.17); X-adenosyl-L-methionine cleaving enzyme or adenosylmethioninase (EC 3.3.1.-); /3-galactosidase (EC 3.2.1.23). This is the second paper in a series; the first paper appeared elsewhere [6]. polymerase was not dependent on the presence of the host termination factor (e) [7]. I n contrast, purified Escherichia coli RNL4 polymerase required the presence of e factor in the reaction mixture to produce RNA species similar in size to those found for early RNA in vivo. Since size alone is only a preliminary criterion on which to base the identity of an RNA species, we have extended these studies through the use of a cell-free protein-synthesizing system programmed by RNA products produced in vivo or in vitro, following their sedimentation through sucrose gradients.The results confirm our previous findings on the involvement of Q factor during the transcription of the T3 genome by the host polymerase and support our origi...

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“…coli RNA polymerase; (b) the results presented in Table 1 were obtained in the presence of rifampicin at a level which completely inhibits E. coli RNA polymerase activity [23].…”

Section: J Dunn W T M C~l I S T E R mentioning

confidence: 96%

Transcription in vitro of T3 DNA by Escherichia coli and T3 RNA Polymerases

Dunn

McAllister

Bautz

1972

European Journal of Biochemistry

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“…The first step involves binding of the enzyme to the DNA template; the subsequent steps are chain initiation, chain elongation, and chain termination. Rifampicin inhibits Escherichia coli RNA polymerase after binding of polymerase to the DNA and before formation of the first internucleotide bonds (1,5,6 into C13CCOOH-insoluble material in 10 min. Antibiotics were dissolved in dimethyl formamide to a concentration of 10 mg/ml as stock solution, and were added to the assay mixture at the desired concentration.…”

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confidence: 99%

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confidence: 99%

Action of Rifamycin Derivatives on RNA Polymerase of Human Leukemic Lymphocytes

Tsai

Saunders

1973

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

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The mechanism of inhibition of human RNA polymerase by four rifamycin derivatives was investigated. Derivative AF/013 (3-formyl rifamycin SV: 0-noctyloxime) with strong hydrophobic side chains prevents the polymerase from binding to DNA and also affects the size of RNA synthesized. Derivative benzyl-2'-methylpyrrolo[3,2-cJ-4-desoxy-rifamycin SV) only affects RNA synthesis when RNA polymerase has been previously incubated with the drug or when the reaction was performed at high salt concentration [0.14 M (NH4)2-S041-Our results suggest that these drugs exert their inhibitory actions by binding to the enzyme instead of DNA.The antibiotic rifampicin inhibits DNA-dependent RNA polymerase obtained from prokaryotic cells, but it has no effect on RNA polymerase isolated from higher organisms (1). One rifamycin derivative, AF/013 (3-formyl rifamycin SV:O-n-octyloxime) synthesized by Lepetit S.p.A., Milan, Italy, can inhibit DNA-dependent RNA polymerases isolated from rat liver (2) and calf thymus (3). This drug inhibits initiation of RNA chains (2, 3). We report studies of the action of rifampicin and four rifamycin derivatives: AF/05 (3-formyl rifamycin SV :O-[diphenylmethyl] oxime), AF/013, PR/19 (3'-acetyl-1'-benzyl-2'-methylpyrrolo[2,3-c] 4-desoxyrifamycin SV), and PR/14 (3'-acetyl-1', 2'-dimethylpyrrolo [3,2-c ]-4-desoxy-rifamycin SV) on DNA-dependent RNA polymerase II (or B) isolated from lymphocytes of patients with chronic lymphocytic leukemia.In vitro RNA synthesis by DNA-dependent RNA polymerase proceeds in a stepwise manner (3, 4). The first step involves binding of the enzyme to the DNA template; the subsequent steps are chain initiation, chain elongation, and chain termination. Rifampicin inhibits Escherichia coli RNA polymerase after binding of polymerase to the DNA and before formation of the first internucleotide bonds (1,5,6

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“…5. The addition of The activity of the ribosome-bound T3 polymerase activity was assayed by the procedure of Dunn et al (8) (17). The inhibition of E. coli RNA polymerase activity by the inhibitor is observed with either T3, T2, or E. coli DNA and, thus, does not depend on the nature of the template.…”

Section: Mechanism Of Inhibitionmentioning

confidence: 99%

An Inhibitory Protein of Escherichia coli RNA Polymerase in Bacteriophage T3-Infected Cells

Mahadik

Dharmgrongartama

Srinivasan

1972

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

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A partially purified protein isolated from bacteriophage T3-infected cells of Escherichia coli B markedly inhibits the activity of E. coli RNA polymerase, slightly inhibits the activity of purified T3 polymerase, and does not inhibit the activity of either core polymerase or the ribosome-bound T3 polymerase. DEAE-cellulose column (2.2 X 20 cm), which was washed with 150 ml of buffer A; the column was eluted with a gradient from 0 to 0.4 M KCl in buffer A. 5-ml fractions were collected, and the fractions containing RNA polymerase activity (0.23-0.3 M KCl) were pooled and diluted 1:1 with buffer C (identical to buffer A, except for 50 mM Tris HCl), and passed through a phosphocellulose column equilibrated with buffer C containing 0.1 M KCl. The inhibitory activity was eluted with the unadsorbed material. This eluate was adjusted to 10 mM MgCl2 and placed on a second DEAEcellulose column (1.8 X 20 cm). The column was eluted with a gradient from 0 to 0.4 M KCl in buffer A containing 10 mM MgCl2. The fractions (0.25-0.3 M KCl) that contained the inhibitory activity were pooled and precipitated with (NH4)2SO4.

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Inhibition of Dna-Dependent Rna Synthesis by Rifamycins

Cited by 151 publications

References 6 publications

Transcription in vitro of T3 DNA by Escherichia coli and T3 RNA Polymerases

Transcription in vitro of T3 DNA by Escherichia coli and T3 RNA Polymerases

Action of Rifamycin Derivatives on RNA Polymerase of Human Leukemic Lymphocytes

An Inhibitory Protein of Escherichia coli RNA Polymerase in Bacteriophage T3-Infected Cells

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