Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.6 (RNA polymerase)
 34,946 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

When Semliki Forest virus ts-4 mutant infected cultures are grown at the permissive temperature (28 degrees C) and shifted to the restrictive temperature (39 degrees C), two different defects in RNA synthesis are manifested: (i) the synthesis of 26S RNA is stopped within 60 min (Saraste et al. 1977) and (ii) the increase in RNA synthesizing activity ceases, in contrast to cultures maintained at 28 degrees C, indicating that no new active RNA polymerase is formed at 39 degrees C. Accumulation of a non-structural precursor protein with an apparent mol. wt. of about 220 000 (ns220) was demonstrated in ts-4 infected cultures shifted to 39 degrees C. NS220 was labelled during short pulses given immediately after release of protein synthesis from hypertonic initiation block, suggesting that genes coding for ns220 are located near the initiation site at the 5'-end of the 42S RNA. The viral specificity of ns220 was shown by its disappearance after a shift to 28 degrees C and by labelling in the presence of sucrose, when no host cell protein synthesis is detectable. The two functional defects can be explained if the polypeptides responsible for the RNA polymerizing activity and that responsible for the synthesis of 26S RNA are components of the same non-structural polyprotein. A mutation in the latter polypeptide which prevents cleavage of the polyprotein would thereby prevent the further formation of active RNA polymerase. If cleavage of the polyprotein has taken place at the permissive temperature, the RNA polymerase would remain active also at 39 degrees C, whereas the polypeptide responsible for 26S RNA synthesis would become inactive due to the mutation.
J Gen Virol 1978 Jun
PMID:Cleavage defect in the non-structural polyprotein of Semliki Forest virus has two separate effects on virus RNA synthesis. 66 Jan 63

Transcriptional complexes formed in vitro with coliphage T5 DNA as template were analyzed by electron microscopy and the number and location of starting sites utilized by E. coli RNA polymerase were determined. Of the 40 promoters characterized in this way, 6 map in the two terminal "pre-early" regions, 29 in the "early" and 5 in the "late" region. The direction of transcription within the different regions determined in this study agrees with earlier findings derived from RNA synthesized in vivo.
Mol Gen Genet 1978 Oct 30
PMID:Electron microscopic analysis of in vitro transcriptional complexes: mapping of promoters of the coliphage T5 genome. 74 95

As an effort to elucidate the control of quality and quantity of the DNA-dependent RNA polymerase in Escherichia coli, the rate of synthesis of the individual subunits was determined during shift-up and -down of nutrients. When the strain B/r grown in a succinate medium was imposed to a shift-up by adding a mixture of glucose and amino acids, rapid rise was observed of the differential rates of the synthesis of alpha, beta and beta' subunits, the constituents of core enzyme, leading to the increase of core polymerase concentration. The differential rates decreased thereafter to the level characteristic of the post-shift rate of cell growth. Compared to the strain B/r, the adaptation was slow in the strain K12 W3350. On the other hand, upon transfer of the strain B/r from a glucose-amino acids medium to a glucose medium lacking amino acids, the differential rate of core polymerase synthesis decreased rapidly and then regained the rate characteristic of the new growth rate. Similar control was also observed on the rate of ribosomal protein synthesis suggesting the coordinate expression of genes for the core polymerase subunits and ribosomal proteins. Thus, the intracellular concentration of RNA polymerase as well as of ribosomes might be one of the most important factors that affect the rate of bacterial growth. The rate of alpha subunit synthesis, however, exhibited little change during the shift-up but a considerable decrease was observed during the shift-down.
Mol Gen Genet 1975 Dec 23
PMID:Biosynthesis of RNA polymerase in Escherichia coli. II. control of RNA polymerase synthesis during nutritional shift up and down. 76 37

Studies on the rate of synthesis of the beta and beta' subunits of RNA polymerase in haploid strains of Escherichia coli K12 containing poorly-suppressed rif degrees am mutations provide conclusive evidence that synthesis of at least these two subunits is regulated.
Mol Gen Genet 1975 Dec 30
PMID:Induction of RNA polymerase synthesis in Escherichia coli. 76 46

E coli RNA polymerase holoenzyme is able to recognize transcription initiation sites on Adenovirus 2 DNA that are functionally indistinguishable from promoters for the enzyme on phage DNAs. The complexes formed between the polymerase and the DNA at these sites can exist in two states-either as I (initiation) complexes, from which rapid RNA chain initiation is not possible, or as RS (rapid starting) "rifampicin resistant" complexes, from which rapid RNA chain initiation can occur. When transcription is limited to that initiated from stable, rifmapicin-resistant pre-initiation complexes, initiation is strictly dependent on the presence of sigma factor; in addition, the frequency of initiation exhibits sigmoidal dependence on the temperature at which pre-initiation complexes are allowed to form, with a transition temperature of 26-28 degrees C. The average half-time for initiation of RNA chains from sites on Ad 2 DNA is shown to be comparable to half-times for initiation of RNA chains from promoters on T7 and lambda DNAs. At saturating levels of enzyme, the half-times are 0.6, 0.9, and 1.6 sec for lambda b2, Ad 2 and T7 DNAs, respectively. The existence of efficient, phage-like promoters for E coli RNA polymerase on Ad 2 DNA suggests to us that such promoters may be closely related functionally and spatially to promoters for mammalian RNA polymerases.
Mol Gen Genet 1976 Jan 16
PMID:I.n vitro transcription of adenovirus 2 DNA. I. Characterization of promoters for E. coli RNA polymerase. 76 51

We estimate that E. coli RNA polymerase is able to form stable, rifampicin-resistant, pre-intiation complexes with Adenovirus 2 DNA at three to six binding sites. The number of RNA chains initiated from such complexes has been determined form the incorporation of gamma-32P-ATP and -GTP at two rifampicin concentrations (7 mug/ml and 24 mug/ml) and after pre-incubation at either 25 or 37 degrees C. The total number of RNA chains initiated ranges from 2.6 per Ad 2 DNA molecule at a rifampicin concentration of 24 mug/ml and pre-incubation temperature of 25 degrees C, to 5.4 per Ad 2 DNA molecule at a rifampicin concentration of 7 mug/ml and pre-incubation temperature of 37 degrees C. Efficient initiation with GTP occurs only after pre-incubation at 37 degrees C whereas initiation with ATP is equally as efficient at either pre-incubation temperature. Promoters for initiation with ATP have been localized to the leftmost 58% of the Ad 2 DNA molecule, defined by the EcoR.RI restriction endonuclease fragment A; promoters for initiation with GTP are located on the remaining 42% of the Ad2 DNA molecule. It is likely that on Adenovirus 2 DNA each RNA chain is initiated from a unique binding site which constitutes a seperate promoter for E. coli RNA polymerase.
Mol Gen Genet 1976 Jan 16
PMID:In vitro transcription of adenovirus 2 DNA. II. Quantification and localization of promoters for E. coli RNA polymerase. 76 52

Temperature-sensitive mutants of Escherichia coli that are unable to grow at high temperature can be obtained among those selected for resistance to streptovaricin or rifampicin at low temperature (Yura et al., 1973). One of these mutants (KY5323) that was supposed to carry a single mutation affecting both rifampicin resistance and temperature sensitivity was further investigated. Using purified RNA polymerase preparations obtained from the mutant and the wild type, it was found that the activity for RNA chain elongation is more sensitive to heat treatment than that for RNA chain initiation or DNA binding, and that the mutant enzyme is significantly more labile than the wild-type enzyme with respect to RNA chain elongation, when heat treatment is carried out at high salt concentration. These results, taken together with those of the enzyme reconstitution experiments, strongly suggest that the beta subunit of the polymerase is directly involved in both RNA chain initiation and elongation reactions. Enzyme reconstitution experiments using isolated subunits derived from the mutant and the wild-type polymerases demonstrate that the alteration of beta subunit is primarily responsible for both rifampicin resistance and thermolability of the mutant enzyme. In addition, the results suggested the apparent alteration of both beta and alpha subunits in this mutant. Extensive transduction experiments provided genetic evidence that are consistent with the view that the strain KY5323 carries a second mutation affecting the beta subunit, beside the primary mutation affecting the beta subunit. The hypothetical beta subunit mutation seems to modify quantitatively the rifampicin resistance caused by the beta subunit mutation.
Mol Gen Genet 1976 Feb 02
PMID:RNA polymerase mutants of Escherichia coli. III. A temperature-sensitive rifampicin-resistant mutant. 76 57

Upon release of rifampicin inhibition of Escherichia coli cells, the initiation of transcription will resume. The sequential resumption of the synthesis of proteins after release of rifampicin inhibition reflects the genetic order and size of the corresponding transcriptional units. We have used this approach to analyze whether the genes for alpha and sigma are on the same transcriptional unit as the genes for beta and beta', employing a method, which allowed us to measure the amounts of RNA polymerase subunits, alpha, beta, beta' and sigma in crude extracts. We have found that the alpha and sigma subunits are synthesized concurrently with the beta subunit in the rifampicin restart experiment, which suggests that the genes for alpha and sigma belongs to different transcriptional units.
Mol Gen Genet 1976 Feb 02
PMID:Biosynthesis of Escherichia coli RNA polymerase subunits upon release of rifampicin inhibition. 76 61

An E. coli mutant rpoA109 unable to support the growth of phage P2 produces DNA-dependent RNA polymerase with an altered alpha subunit. Histidine is substituted for leucine in one tryptic peptide from the mutant alpha subunit. The existence of only one rpoA gene within the E. coli chromosome is indicated.
Mol Gen Genet 1976 Apr 23
PMID:Identification of a mutation within the structural gene for the a subunit of DNA-dependent RNA polymerase of E. coli. 77 6

We describe a bacterial RNA polymerase mutation, rif 501, which confers rifampicin resistance and thermosensitivity to E. coli K 12. The purified RNA polymerase enzyme from rif 501 bacteria shows increased heatsensitivity in vitro at 51 degrees C. However, in vivo, at 42 degrees C the non-permissive temperature, mutant bacteria continue to grow and to synthesize RNA for 90 min. On a lawn of the mutant bacteria, at 40-41 degrees C, phage lambda forms clear plaques (LycA phenotype); this is probably due to an enhancement of cro function; we surmise that at 42 degrees C the transcription originating from the pR (but not from the pL) promoter on the lamdba genome becomes N-independent and less sensitive to the absence of the cro product. We discuss the possibility that both the N and cro proteins of phage lambda interact directly with the bacterial RNA polymerase. These observations indicate that the loss of viability of the rif 501 mutant at the restrictive temperature is not a consequence of an immediate inactivation of RNA polymerase; rather we feel it is due to a modification of the activity of RNA polymerase, leading to a disruption of the cellular regulation.
Mol Gen Genet 1976 Apr 23
PMID:A bacterial RNA polymerase mutant that renders lambda growth independent of the N and cro functions at 42 degrees C. 77 9


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