Regulation of Metabolic Breakdown and Synthesis of Messenger RNA in Bacteria

Gros, François; Dubert, J M; Tissières, A.; Bourgeois, Suzanne; Michelson, M; Soffer, Richard L.; Legault, Lucienne

doi:10.1101/sqb.1963.028.01.045

Cited by 45 publications

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“…The difference in the half-life of alkaline phosphatase mRNA between B. subtilis (ti = 1.6 min; 6) and E. coli could be either species-specific or due to differences in the experimental conditions (11). Basically, however, the same nonsense strain was used in our experiments and in those reported by Rosen (24), and by Edlin and Maal0e (5); we believe, therefore, that the shorter life span of the mRNA molecule obtained in our case is due to the fact that actinomycin D was used for the termination of RNA synthesis, whereas Rosen and Edlin and Maal0e used excess uracil for chasing the 5FU.…”

Section: Resultsmentioning

confidence: 99%

Decay of Normal and 5-Fluorouracil-Substituted Messenger Ribonucleic Acid of Alkaline Phosphatase in Escherichia coli

Yagil

Silberstein

1969

J Bacteriol

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The decay of alkaline phosphatase messenger ribonucleic acid (mRNA) in Escherichia coli was studied in cells sensitized to actinomycin D by ethylenediaminetetra-acetate treatment. It was found that in the wild-type strain (K-10) as well as in a nonsense mutant (S26C200), which is phenotypically reversible by treatment with 5-fluorouracil, mRNA decays with a half-life of 2.0 to 2.5 min. Similarly, in a 3-min pulse of labeled 5-fluorouracil, 70% of the incorporated analogue is contained in the labile RNA fraction decomposing with a half-life of 1.8 min.

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Section: Resultsmentioning

confidence: 99%

Decay of Normal and 5-Fluorouracil-Substituted Messenger Ribonucleic Acid of Alkaline Phosphatase in Escherichia coli

Yagil

Silberstein

1969

J Bacteriol

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“…That various polynucleotides can inhibit RNA polymerase is well established [S, 11,43,44]. It has been reported that the inhibition is reduced a t higher ionic strength [45].…”

Section: )Ooimentioning

confidence: 99%

Influence of Salts on RNA Synthesis by DNA‐Dependent RNA‐Polymerase from Escherichia coli

Fuchs

Millette

Zillig

et al. 1967

European Journal of Biochemistry

191

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The effect of salt concentration on RNA synthesis by DNA-dependent RNA-polymerase from Escherichia coli has been investigated. For a double stranded template the following influences are observed: (1) Free enzyme and enzyme bound to DNA, but not synthesizing RNA, are inhibited above an ionic strength of about 0.15. (2) The rate of RNA-synthesis, by those enzyme molecules which have started RNA chains, is stimulated by increasing the salt concentration up to an optimum. Magnesium salts cause a larger stimulation than monovalent cations. However, the optimal effect always occurs at an ionic strength of about 0.36. (3) This stimulatory effect is distinguished from the cofactor function which requires only low concentrations of divalent cations (Mg2+, Co2+, Mn2+). (4) RNA synthesis stops at low ionic strength (< 0.2) due to a first order inactivation of the synthesizing enzyme. A t high ionic strength synthesis continues over many hours a t a slowly decreasing rate. Raising the ionic strength after RNA synthesis has stopped at low ionic strength causes an immediate resumption of synthesis. Mg2+ and spermidine are most effective in this action. However, Co2+ and Mn2+ are unable to reactivate the enzyme once it has stopped.With single stranded DNA as template neither a first order inactivation of synthesizing enzyme a t low ionic strength nor a stimulation of RNA synthesis by monovalent> salts is observed. The effect of divalent cations on enzyme activity can be due to their cofactor function.The DNA-dependent RNA-polymerase from Escherichia coli is strongly influenced by changes in the ionic strength. At low ionic strength (between 0.02 and 0.12) it has been shown to exist in a form sedimenting a t 24s [l-51; a t high ionic strength the enzyme reversibly dissociates into subunits of 13s [2,4-61. At low salt concentration the polymerase forms a complex with DNA [1,7--12j which by raising the ionic strength is dissociated into free DNA and the i3S form of the enzyme [Z, 13-15].The polymerization process itself has been reported to be inhibited only after some time of synthesis at high ionic strength [5,13]. However, the effect of ionic strength on this phase of the reaction has not been extensively investigated.This communication presents a detailed analysis of the effect of ionic strength on the kinetics of RNAsynthesis with double and single-stranded DNA. It is shown that increasing the ionic strength causes Non-Standard Abbreviations. PPO, 2,5 diphenyloxazol; POPOP, 1,4-bis-2-(4 methyl4 phenoxazoly1)-bcnzene; TMA, buffer, containing: 0.01 M Tris acetate, 0.022 M NH,CI, 0.01 M magnesium acetate, 1 mM 2-mercaptoethanol, pH 7.3; TCA, trichloroacetic acid.Enzymes. DNA-dependent RNA-polymerase or nucleosidetripliosphate : RNA nucleotidyltransferase (EC 2.7.7.6) ; Ribonuclease I (EC 2.7.7.17); Pyruvate kinase (EC 2.7.1.40). a striking stimulation of both the rate and extent of RNA synthesis. This eliminates the typical plateau in the kinetics [l6-211 and allows RNA-synthesis t o continue over many hours. Moreover, synthes...

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“…Earlier studies by Niyogi and Stevens (9) with holo RNA polymerase and synthetic polyribonucleotide templates indicated that only complementary oligoribonucleotides with a free 3'-hydroxyl group are stimulatory and preferentially incorporated into the 5'phosphomonoester chain end of the product, hence acting as chain initiators. Gros et al (10), using Escherichia coli DNA as template and normal substrate concentrations, observed small stimulations by various oligoribonucleotides. Downey et al (11) reported that certain dinucleoside monophosphates stimulate chain initiation and the overall rate of RNA synthesis at low substrate concentrations in reactions using either T4 DNA and holoenzyme or T5 DNA with core enzyme or holoenzyme.…”

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RNA Initiation with Dinucleoside Monophosphates during Transcription of Bacteriophage T4 DNA with RNA Polymerase of Escherichia coli

Hoffman

Niyogi

1973

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

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The effects of dinucleoside monophosphates on the transcription of phage T4 DNA by E. coli RNA polymerase have been examined at various concentrations of the sigma subunit and extremely low concentration of ribonucleoside triphosphate. The following conclusions were reached: (i) Labeled specific dinucleoside monophosphates are incorporated as chain initiators. (ii) When the ratio of sigma factor to core enzyme is small, there is a general stimulation by most 5'-guanosyl dinucleoside monophosphates. (iii) When the ratio is increased or holoenzyme is present, ApU, CpA, UpA, and GpU are the most effective stimulators. (iv) At high concentrations of sigma factor, only certain adenosine-containing dinucleoside monophosphates (ApU, CpA, UpA, and ApA) stimulate the reaction. (v) Competition hybridization studies indicate that the RNAs stimulated by dinucleoside monophosphates (ApU, CpA, UpA, and GpU) are of the T4 "early" type. (vi) Studies involving both combinations of stimulatory dinucleoside monophosphates and competitive effects of these compounds on chain initiation by ATP and GTP suggest that the stimulatory dinucleoside monophosphates act as chain initiators and may recognize part of a continuous sequence in a promoter region. Studies based on the incorporation of 3H-labeled stimulatory dinucleoside monophosphates support the above conclusions.The discovery of the sigma (a) subunit of bacterial RNA polymerase added strength to the concept of positive control of transcription in vitro (1-3). a-Factor is a necessary requirement for accurate initiation of RNA synthesis by core RNA polymerase in vitro on certain phage DNA templates (4-7). Our studies with well-defined templates have shown that the stimulatory effect of o-factor depends on both the base composition and secondary structure of the template, as well as the concentration of ribonucleoside triphosphate, and that the same type of stimulatory effect is obtained with oligoribonucleotides complementary to the templates (8). Earlier studies by Niyogi and Stevens (9) with holo RNA polymerase and synthetic polyribonucleotide templates indicated that only complementary oligoribonucleotides with a free 3'-hydroxyl group are stimulatory and preferentially incorporated into the 5'phosphomonoester chain end of the product, hence acting as chain initiators. Gros et al. (10)

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Regulation of Metabolic Breakdown and Synthesis of Messenger RNA in Bacteria

Cited by 45 publications

References 0 publications

Decay of Normal and 5-Fluorouracil-Substituted Messenger Ribonucleic Acid of Alkaline Phosphatase in Escherichia coli

Decay of Normal and 5-Fluorouracil-Substituted Messenger Ribonucleic Acid of Alkaline Phosphatase in Escherichia coli

Influence of Salts on RNA Synthesis by DNA‐Dependent RNA‐Polymerase from Escherichia coli

RNA Initiation with Dinucleoside Monophosphates during Transcription of Bacteriophage T4 DNA with RNA Polymerase of Escherichia coli

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