Basis of Actinomycin Action, I. Dna Binding and Inhibition of Rna-Polymerase Synthetic Reactions by Actinomycin

Goldberg, Irving H.; Rabinowitz, Murray; Reich, E.

doi:10.1073/pnas.48.12.2094

Cited by 331 publications

(90 citation statements)

References 15 publications

(3 reference statements)

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“…Inhibition probably results from impaired RNA synthesis since low concentrations of actinomycin D selectively suppresses DNA-directed RNA synthesis (Goldberg, Rabinowitz, and Reich 1962). DNA synthesis is affected by actinomycin D only at much higher concentrations.…”

Section: Introduotionmentioning

confidence: 99%

Effect of Actinomycin D and 2-Thiouracil on Floral Induction and Nucleic Acid Synthesis in the Bud in Chenopodium Amaranticolor

Watson¹,

Matthews²

1966

Aust. Jnl. Of Bio. Sci.

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SummaryActinomycin D and 2-thiouracil inhibit floral initiation in O. amaranticolor when applied to plants before the end of the inductive dark period. Actinomycin D does not suppress floral differentiation if it is applied to plants more than 48 hr after the inductive dark cycle, whereas 2-thiouracil retains its ability to interfere with floral differentiation when applied to plants several days after the inductive dark period.A new, 32P-labelled, ribonuclease-resistant RNA component was detected in apical buds following floral induction. Fractionation of 32P-labelled RNA isolated from induced apical buds on columns of methylated albumin on kieselguhr showed this to be associated with the DNA probably as a DNA-RNA hybrid. This RNA component was not detected in non-induced apical buds.Actinomycin D and 2-thiouracil decrease the incorporation of [32P]ortho_ phosphate into apical bud RNA in induced apical buds. Both inhibitors suppressed the appearance of the 32P-label1ed, ribonuclease-resistant RNA component found in non-treated, induced apical buds. DNA-RNA hybrids may be needed for the synthesis of messenger RNA needed for floral bud differentiation. L INTRODUOTIONNucleic acids are undoubtedly closely involved in the processes that are initiated in vegetative apices following photoperiodic induction of flowering. A number of inhibitors of nucleic acid synthesis have been used in attempts to obtain more detailed information concerning these processes. These include fluoropyrimidines, actinomycin D, and 2-thiouracil. 5-Fluorouracil inhibits floral induction in Xanthium (Bonner and Zeevaart 1962) and in Pharbitis (Zeevaart 1962) when applied at the beginning of the inductive dark period. In both of these short-day plants 5-fluorouracil inhibits the synthesis of DNA and RNA in the buds. 5-Fluorodeoxyuridine inhibits floral induction in Pharbitis and XantMum (Zeevaart 1962) and its action is more specific than 5-fluorouracil (Harbers, Chaudhuri, and Heidelberger 1959). Studies with 5-fluorodeoxyuridine suggest that DNA synthesis in the apical bud is essential for floral induction.Actinomycin D inhibits floral induction in Lolium (Evans 1964) and in Pharbitis (Galum, Gresset, and Keynam 1964). Inhibition probably results from impaired RNA synthesis since low concentrations of actinomycin D selectively suppresses

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Section: Introduotionmentioning

confidence: 99%

Effect of Actinomycin D and 2-Thiouracil on Floral Induction and Nucleic Acid Synthesis in the Bud in Chenopodium Amaranticolor

Watson¹,

Matthews²

1966

Aust. Jnl. Of Bio. Sci.

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“…DNA synthesis is not affected materially by low concentrations of AMD (4). Actinomycin apparently binds to DNA (5), probably specifically with guanine (6,7), in bringing about its biological effects. These properties of AMD have led us to study its effects on cell differentiation in vitro using the technique of small aggregate cultures of amphibian embryonic cells developed in this laboratory (8).…”

mentioning

confidence: 99%

The Effects of Actinomycin D on the Ultrastructure of the Nucleus of the Amphibian Embryonic Cell

Jones¹,

Elsdale²

1964

The Journal of Cell Biology

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Observations have been made of ultrastructural modifications induced in the nuclei of differentiating amphibian embryonic cells cultured in the presence of Actinomycin D. Of particular interest are regions within the nucleus (regions otherwise rather empty) containing loose groupings of uniform threads having a diameter of around 200 A. These threads have been observed in continuous lengths up to 0.5 #, and appear to be composed of subfilaments.

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“…B) The inakility of ithe plianit enzyme to utilize RNA or synthetic oligoribonucleotide templates (42) is in contrast with their utiilization by microbial enzymes (16,22,30,40). C) The formation of homopolymers in the presence of he!ated DNA is catalyzed by the microbial enzymes (6,40), wherea,s the formation of a homopolymer is not detected with the maize polymerase.…”

Section: Methodsmentioning

confidence: 99%

Template Requirement of Maize RNA Polymerase

Stout

Mans

1968

Plant Physiol.

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A bstract. Maize RNA polymerase utilizes heated deoxyribonucleic acid more effectively than native deoxyribonucleic acid as a template for ribonucleic acid synthesis. A ribenucleic acid-deoxyribonucleic acid hybrid accumulates in the presence of 'heated deoxyribonucleic acid. The amount of product formed with either native or heat-denatured deoxyribonucleic acid does not exceed the amount of deoxyribonucleic acid added as template.The itemplate requirement of several RNA polymerases ('Nucleoside-Htriphosphate: RNA nudleotidyl transferase, EC 2.7.7.6), isolated and purified from microorganism's (5,7,11,41 ) anid mamtmals ( 13, 32), is best satisfied with native DNA. Heat denatured, calf thymus DNA actually inhibits the polymerase isolated from bovine lymiphosarcoma tissue (12). In characterizing the partially purified enzyme from maize seedllings we have shown that, lat relatively high levels, native or denatured DNA from several species satisfies the template requirement of the maize enzyme (42). The data presented here demonstrate thait at rate limiting concentrations, denatured DNA from maize seed1lings or calf thym'us is 8 times more effective than native DNA in satisfying the template requirement. Furthermore, an RNA-DNA hybrid, analogous to that formed by Azotobacter vinelandii RNA polymerase (44), accutmulates u'pon incuLbation of the enzyme with 'sub'stra'te and denatured DNA. The hybrid is not found w'hen native DNA is used as template. Materials and MethodsEnzyme Preparationt and Assay. Soluble RNA polymerase was isolated f'rom 5-day-old maize seedlings as described previously (42). The am'monium sullfate precipitate was desalited on Sephadex G-50 ra,ther than by di,a)lysis prior to column chromatography on diethylaminoethyl-( DEAE) cellu'lose.The enzyme had a specific activity of 1870. Specific activity is def'ined as rnLmoles of AMP incorporated in 10 minutes at 300 per milligram prote'in.

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Basis of Actinomycin Action, I. Dna Binding and Inhibition of Rna-Polymerase Synthetic Reactions by Actinomycin

Abstract: Actinomycin inhibits DNA-dependent RNA synthesis of intact cells' and enzymes from mammalian2 and bacterial' sources but not the growth of RNA-viruses in

Cited by 331 publications

References 15 publications

Effect of Actinomycin D and 2-Thiouracil on Floral Induction and Nucleic Acid Synthesis in the Bud in Chenopodium Amaranticolor

Effect of Actinomycin D and 2-Thiouracil on Floral Induction and Nucleic Acid Synthesis in the Bud in Chenopodium Amaranticolor

The Effects of Actinomycin D on the Ultrastructure of the Nucleus of the Amphibian Embryonic Cell

Template Requirement of Maize RNA Polymerase

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