Primer RNA Synthesis by E. coli RNA Polymerase on the SSB-coated 229-nt ssi Signal of Lactococcal Plasmid pGKV21

9-Aminoacridine carboxamide derivatives studied here form with DNA intercalative complexes which differ in the kinetics of dissociation. Inhibition of total RNA synthesis catalyzed by phage T7 and Escherichia coli DNA-dependent RNA polymerases correlates with the formation of slowly dissociating acridine-DNA complex of time constant of 0.4-2.3 s. Their effect on RNA synthesis is compared with other ligands which form with DNA stable complexes of different steric properties. T7 RNA polymerase is more sensitive to distamycin A and netropsin than the E. coli enzyme while less sensitive to actinomycin D. Actinomycin induces terminations in the transcript synthesized by T7 RNA polymerase. Despite low dissociation rates of DNA complexes with acridines and pyrrole antibiotics no drug dependent terminations are observed with these ligands.

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Translocation of DNA-Dependent E. coli RNA Polymerase During RNA Synthesis

Mechanisms of Transcription - Nucleic Acids and Molecular Biology ◽

10.1007/978-3-642-60691-5_11 ◽

1997 ◽

pp. 151-177 ◽

Cited By ~ 4

Author(s):

H. Heumann ◽

E. Zaychikov ◽

L. Denissova ◽

T. Hermann

Keyword(s):

Rna Polymerase ◽

Rna Synthesis ◽

E Coli

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Transcription of bacteriophage T4 deoxyribonucleic acid in vitro

Biochemical Journal ◽

10.1042/bj1150353 ◽

1969 ◽

Vol 115 (3) ◽

pp. 353-361 ◽

Cited By ~ 13

Author(s):

John O. Bishop ◽

Forbes W. Robertson

Keyword(s):

Rna Polymerase ◽

Messenger Rna ◽

Rna Synthesis ◽

Deoxyribonucleic Acid ◽

Bacteriophage T4 ◽

Rna Sequences ◽

Experimental Conditions ◽

E Coli ◽

New Methods

1. RNA was synthesized in vitro from a template of bacteriophage T4 DNA, in the presence of Mn2+. A comparison was made of the RNA synthesized by purified RNA polymerase from two sources, Micrococcus lysodeikticus and Escherichia coli; these are referred to as Micrococcus cRNA and E. coli cRNA respectively (where cRNA indicates RNA synthesized in vitro by using purified RNA polymerase and a DNA primer). 2. Both types of RNA were self-complementary as judged by resistance to digestion with ribonuclease after self-annealing, Micrococcus cRNA being more self-complementary (40%) than was E. coli cRNA (30%). The cRNA was found to be much less self-complementary if Mg2+ was present during RNA synthesis instead of Mn2+. 3. Micrococcus cRNA hybridized with a larger part of bacteriophage T4 DNA than did E. coli cRNA. The E. coli cRNA competed with only part (70%) of the Micrococcus cRNA in hybridization-competition experiments. It is concluded that more sequences of bacteriophage T4 DNA are transcribed by Micrococcus polymerase than by E. coli polymerase. 4. The RNA sequences synthesized by Micrococcus RNA polymerase but not by E. coli RNA polymerase are shown by hybridization competition to compete with specifically late bacteriophage T4 messenger RNA sequences. The relevance of this finding to the control of transcription is discussed. 5. In an Appendix, new methods are described for the analysis of hybridization-saturation and -competition experiments. Particular attention is paid to the effects produced if different RNA sequences are present at different relative concentrations. 6. By using cRNA isolated from an enzymically synthesized DNA–RNA hybrid, it is estimated that, of the DNA that is complementary to cRNA, only about half can become hybridized with cRNA under the experimental conditions used.

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The properties of ATP-analogs in initiation of RNA synthesis catalyzed by RNA polymerase from E. coli

Nucleic Acids Research ◽

10.1093/nar/9.10.2397 ◽

1981 ◽

Vol 9 (10) ◽

pp. 2397-2410 ◽

Cited By ~ 3

Author(s):

W.J. Smagowicz ◽

K.H. Scheit

Keyword(s):

Rna Polymerase ◽

Rna Synthesis ◽

E Coli ◽

Atp Analogs

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Effect of multiple copies ofrpoBCon the rate of RNA synthesis inEscherichia coli

Genetics Research ◽

10.1017/s0016672300024770 ◽

1986 ◽

Vol 48 (2) ◽

pp. 61-64 ◽

Cited By ~ 2

Author(s):

Elena C. Guzman ◽

Alfonso Jimenez-Sanchez

Keyword(s):

Dna Replication ◽

Rna Polymerase ◽

Rna Synthesis ◽

Synthetic Activity ◽

Gene Products ◽

Replication Forks ◽

E Coli ◽

Multiple Copies ◽

Autogenous Regulation ◽

Rate Of Movement

SummaryThe cloning of therpoBandrpoCgenes in a high copy number vector inE. coliincreased the amount of the encoded gene products, the β and β′ subunits of RNA polymerase. However, this unexpectedly caused a 30–50% decrease in RNA synthetic activity which alternatively induced a reduction of growth rate and enlargement of cell size, and decreased the DNA replication time. The results can be explained by autogenous regulation of the RNA polymerase genes by the ββ′ subunits. A relation between the decrease in number of transcription units and the observed higher rate of movement of DNA replication forks is discussed.

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Dual role of the σ factor in primer RNA synthesis by bacterial RNA polymerase

FEBS Letters ◽

10.1002/1873-3468.13312 ◽

2018 ◽

Vol 593 (3) ◽

pp. 361-368

Author(s):

Daria Esyunina ◽

Danil Pupov ◽

Andrey Kulbachinskiy

Keyword(s):

Rna Polymerase ◽

Rna Synthesis ◽

Dual Role ◽

Bacterial Rna ◽

Σ Factor ◽

Primer Rna

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Antimicrobial Properties and Mode of Action of the Pyrrothine Holomycin

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.2.532-539.2001 ◽

2001 ◽

Vol 45 (2) ◽

pp. 532-539 ◽

Cited By ~ 71

Author(s):

Brunello Oliva ◽

Alexander O'Neill ◽

Jenny M. Wilson ◽

Peter J. O'Hanlon ◽

Ian Chopra

Keyword(s):

Rna Polymerase ◽

Rna Synthesis ◽

Antimicrobial Properties ◽

Stringent Response ◽

Active Species ◽

Chain Elongation ◽

Whole Cells ◽

E Coli ◽

Eukaryotic Microorganisms ◽

Kinetics Of

ABSTRACT Holomycin, a member of the pyrrothine class of antibiotics, displayed broad-spectrum antibacterial activity, inhibiting a variety of gram-positive and gram-negative bacteria, with the exception ofEnterobacter cloacae, Morganella morganii, andPseudomonas aeruginosa. The antibiotic lacked activity against the eukaryotic microorganisms Saccharomyces cerevisiae and Candida kefyr. Holomycin exhibited a bacteriostatic response against Escherichia coli that was associated with rapid inhibition of RNA synthesis in whole cells. Inhibition of RNA synthesis could have been a secondary consequence of inhibiting tRNA aminoacylation, thereby inducing the stringent response. However, the levels of inhibition of RNA synthesis by holomycin were similar in a stringent and relaxed pair of E. coli strains that were isogenic except for the deletion of therelA gene. This suggests that inhibition of RNA synthesis by holomycin could reflect direct inhibition of DNA-dependent RNA polymerase. Examination of the effects of holomycin on the kinetics of the appearance of β-galactosidase in induced E. colicells was also consistent with inhibition of RNA polymerase at the level of RNA chain elongation. However, holomycin only weakly inhibitedE. coli RNA polymerase in assays using synthetic poly(dA-dT) and plasmid templates. Furthermore, inhibition of RNA polymerase was observed only at holomycin concentrations in excess of those required to inhibit the growth of E. coli. It is possible that holomycin is a prodrug, requiring conversion in the cell to an active species that inhibits RNA polymerase.

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NusG inhibits RNA polymerase backtracking by stabilizing the minimal transcription bubble

eLife ◽

10.7554/elife.18096 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 36

Author(s):

Matti Turtola ◽

Georgiy A Belogurov

Keyword(s):

Rna Polymerase ◽

Rna Synthesis ◽

Transcription Elongation ◽

Single Nucleotide ◽

E Coli ◽

Transcription Bubble ◽

Nucleotide Addition ◽

Nascent Rna ◽

Comprehensive Mapping

Universally conserved factors from NusG family bind at the upstream fork junction of transcription elongation complexes and modulate RNA synthesis in response to translation, processing, and folding of the nascent RNA. Escherichia coli NusG enhances transcription elongation in vitro by a poorly understood mechanism. Here we report that E. coli NusG slows Gre factor-stimulated cleavage of the nascent RNA, but does not measurably change the rates of single nucleotide addition and translocation by a non-paused RNA polymerase. We demonstrate that NusG slows RNA cleavage by inhibiting backtracking. This activity is abolished by mismatches in the upstream DNA and is independent of the gate and rudder loops, but is partially dependent on the lid loop. Our comprehensive mapping of the upstream fork junction by base analogue fluorescence and nucleic acids crosslinking suggests that NusG inhibits backtracking by stabilizing the minimal transcription bubble.

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