Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

RNA polymerase was isolated from Escherichia coli and Serratia marcescens. The subunits of both enzymes were separated by electrophoresis on cellulose acetate sheets in the presence of urea. Under conditions favouring reconstitution of the RNA polymerases, stoichiometric amounts of the subunits were allowed to interact. Active hybrid enzymes were formed if corresponding subunits of both enzymes were mutually exchanged. The analysis of the RNA products synthesized showed that the reconstituted enzymes are able to recognize the promoters for transcription and the termination signals on the DNA template. The transcription products can serve as messengers for cell-free protein synthesis.
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PMID:Functional hybrid enzymes reconstituted from Escherichia coli and Serratia marcescens RNA polymerase subunits. 77 Jan 71

5-Formyl-1-(alpha-D-ribofuranosyl)uracil 5'=triphosphate has been used to affinity label E. coli DNA-dependent RNA polymerase. It is a noncompetitive inhibitor of the enzyme with Ki=0.54 mM. A short preincubation of the enzyme and alpha-fo5UTP is required to achieve maximum inhibition, and the entent of the inhibition is dependent upon the alpha-fo5UTP concentration. When a preincubation mixture of alpha-fo5UTP/enzyme is diluted, the enzyme regains activity with time showing that the inhibition is reversible, presumably occurring by Schiff base formation between an amino group on the enzyme and the formyl group. Upon sodium borohydride reduction of an enzyme/alpha-fo5UTP preincubation mixture the enzyme is irreversibly inhibited. alpha-fo5UTP is more effective in inhibiting the enzyme than alpha-fo5U, and the inhibition is decreased by the presence of ATP, UTP, or GTP in the preincubation mixture, suggesting that inhibition is occurring at a triphosphate binding site. The stoichiometry of binding of alpha-fo5UTP to the enzyme was determined using the gamma-32P-labeled derivative. After a 20-s preincubation of enzyme/alpha-fo5UTP followed by NaBH4 reduction the stoichiometry of binding was 1.1:1 (alpha-fo5UTP bound: inactivated enzyme), and this rose to 2.42:1 after a 10-min preincubation. After a 20-s preincubation the [gamma-32P]-alpha-fo5UTP was shown to be located on the beta subunit of RNA polymerase by cellulose acetate electrophoresis in 6 M urea.
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PMID:Affinity labeling of Escherichia coli DNA-dependent RNA polymerase with 5-formyl-l-(alpha-D-ribofuranosyl)uracil 5'-triphosphate. 77 15

RNA polymerase of E. coli (EC 2.7.7.6) is able to bind certain oligoribonucleotides with the length greater than or equal to 5 from the corresponding isoplith mixtures (Knorre V.L., Vasilenko S.V., Salganik R.I., FEBS Letters 30, 229, 1973). It has been shown in this study that all pentaribonucleotides able to be bound by RNA polymerase can be extracted from the random mixture by the enzyme saturation procedure. Loosely and tightly bound pentaribonucleotides subfractions were isolated and each was separated by chromatography into 3-4 isopliths. Blocking of the enzyme SH groups by p-chloromercurium benzoate (10(-3) M) and denaturation by urea (6.3 M) prevent formation of the enzyme-pentaribonucleotides complexes. Complexes are destroyed by heat denaturation. Removal of sigma-subunit does not influence the enzyme capacity for pentaribonucleotides binding.
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PMID:[Conditions for specific oligoribonucleotide binding with E. coli RNA-polymerase]. 78 22

Chromosomal non-histone proteins are obtained from nuclei of two types of pigeon erythroid cells: erythroblasts (cells active in RNA synthesis) and erythrocytes (cells with repressed RNA synthesis). They are well soluble in solutions of low ionic strength. Electrophoretic separation of the obtained non-histone proteins in polyacrylamide gels with urea and SDS shows the presence of qualitative differences in the pattern of non-histone proteins of chromatine from erythroblasts and erythrocytes. By electrophoresis in urea some protein bands of non-histone proteins of chromatine from erythroblasts were found which disappear with the aging of cells. At the same time two protein fractions were observed in chromatine from erythrocytes which were absent in that of erythroblasts. Disappearance of some high molecular weight protein fractions from erythrocyte chromatine as compared to erythroblasts was observed by separation of the non-histone proteins in the presence of SDS. These fractions of the non-histone proteins disappearing during aging of cells are well extractable from erythroblast chromatine by 0.35 M NaCl solution. In the in vitro system with E. coli RNA polymerase addition of non-histone proteins of chromatine from erythroblasts to chromatine from erythrocytes increases RNA synthesis 2--3 times. At the same time addition of non-histone proteins from erythrocytes is either without any influence on this process or somewhat inhibiting.
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PMID:[Comparative investigation of the non-histone proteins of chromatin from pigeon erythroblasts and erythrocytes]. 80 71

Nucleic acid-free extracts of Escherichia coli have been analyzed by chromatography on columns of cellulose, to which poly(A), poly(U), or poly(C) have been attached by ultraviolet irradiation. Proteins are released from the columns by stepwise elution with increasingly higher concentrations of salt, followed by washing with urea to remove very tightly bound molecules. The pattern of protein elution is reproducibly different for each of the homopolymer RNA-cellulose columns used: some proteins bind very tightly to one column, but poorly to others. Analysis by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis, by immunological cross-reactivity in double diffusion tests, and by enzymological assays, has allowed the identification of a number of these proteins. The RNA polymerase core enzyme binds to poly(C)- and to poly(U)-cellulose columns, and can be purified to 20 to 30 percent homogeneity in a single step. Ribosomal protein S1 and the termination factor rho bind very tightly to poly(C)-cellulose, and both can be purified to homogeneity rapidly, in much higher yields than previously reported. Poly(A)-cellulose chromatography allows the isolation of large amounts of an 80,000 molecular weight protein having an as yet unassigned cellular function. The host factor required for RNA phage Qbeta RNA replication in vitro can also be obtained from poly(A)-cellulose, and chromatography of extracts of phage Qbeta-infected E. coli on RNA-cellulose columns results in very rapid isolation of the Qbeta replicase enzyme. Homopolymer RNA-cellulose chromatography thus appears to be a simple, general technique, useful for the efficient isolation of a variety of RNA-binding proteins.
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PMID:Isolation of bacterial and phage proteins by homopolymer RNA-cellulose chromatography. 80 80

The DNA-dependent RNA polymerase of the blud-green alga Anacystis nidulans was reconstituted from its isolated subunits in the absence of urea. Applying this technique the kinetics and the subunit requirements of the reconstitution process were analyzed. The results reveal differences with respect to the reconstitution of Escherichia coli polymerase. Reconstitution proceeds much more slowly in the case of the A. nidulans enzyme. Reconstitution here is absolutely dependent on the presence of the subunit sigma. On the other hand, the largest of the subunits of Mr=190000 can be fully substituted by a specific degradation product of this subunit of Mr=175000. Heterologous reconstitution between subunits of E. coli and A. nidulans polymerase does not result in active enzyme hybrids, showing a divergent evolution of the structure of this enzyme in these procaryotic organisms.
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PMID:The reconstitution of Anacystis nidulans DNA-dependent RNA polymerase from its isolated subunits. 81

Partially purified rat liver RNA polymerase I chromatographed on ribosomal RNA-Sepharose loses most (96%) of its activity assayed on native calf-thymus DNA templates, but loses little (8%) of its activity assayed on poly(deoxycytidylic acid) template. Polymerase I is not stimulated by polymerase II protein factor, or by bovine serum albumin. However, it is stimulated by histones, polylysine, and spermine. Addition of a protein fraction eluted by high ionic strength from the rRNA-Sepharose also restores activity on native calf-thymus DNA. Further purification yields a fraction containing two proteins of 11 000 and 12 000 molecular weight. Both proteins are distinct from histones by electrophoresis in sodium dodecyl sulfate and in acid urea. Both proteins are basic, insensitive to heat, bind to DNA, and stimulate polymerase I activity. The degree of stimulation of polymerase I is dependent upon both the enzyme/DNA and the factor/DNA ratio. The protein factors also stimulate polymerase II activity about half as effectively as polymerase I.
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PMID:A protein cofactor that stimulates the activity of DNA-dependent RNA polymerase I on double-stranded DNA. 85 26

We have previously presented a rapid, high yield method for the large scale purification of homogeneous RNA polymerase II from wheat germ (Jendrisak, J.J., and Burgess, R.R.(1975), Biochemistry 14, 4639), and we now report a detailed study of its subunit structure. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicates that polypeptides with molecular weights of 220 000, 140 000, 40 000, 27 000, 25 000, 21 000, 20 000, 17 800, 17 000, 16 500, 16 000, and approximately 14 000 are associated with the enzyme. Two-dimensional polyacrylamide gel electrophoresis, by which the subunits were separated in the first dimension in the presence of 8 M urea at pH 8.7 and in the second dimension in the presence of 0.1% sodium dodecyl sulfate, indicates that the 40 000 molecular weight component is composed of two nearly identical polypeptides and that the low molecular weight components (smaller than or equal to 40 000) are acidic proteins except for the 25 000 molecular weight polypeptide.
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PMID:Studies of the subunit structure of wheat germ ribonucleic acid polymerase II. 85 83

Addition of urea to an uninduced culture of Saccharomyces at 22 C results in appearance of allophanate hydrolase activity after a lag of 12 min. We have previously demonstrated that both ribonucleic acid (RNA) and protein synthesis are needed for this induction to occur. To elucidate the time intervals occupied by known processes involved in induction, temperature-sensitive mutants defective in messenger RNA transport from nucleus to cytoplasm (rna1) and in protein synthesis initiation (prt1) were employed along with an RNA polymerase inhibitor in experiments that measure cumulative synthetic capacity to produce allophanate hydrolase. These measurements identify the time within the lag period at which each of the above processes is completed. We observed that RNA synthesis, rna1 gene product function, and protein synthesis initiation are completed at 1 to 1.5, 4, and 9 to 10 min, respectively.
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PMID:Sequence of molecular events involved in induction of allophanate hydrolase. 94 80

The nonhistone proteins of Ehrlich ascites tumor chromatin have been separated into a loosely bound and two tightly bound protein fractions by sequential extraction of chromatin with 0.35 M NaCl and 2 M NaCl:5 M urea. The nonhistone proteins thus obtained were examined for their chemical composition and distribution of DNA polymerase, RNA polymerase, and protein kinase activities. In addition, the effect of these nonhistone proteins on transcription of DNA in vitro has been determined. The results indicate that these nonhistone proteins, fractionated on the basis of their extractability, exhibit varied compositional characteristics and play different functional roles in the synthesis of DNA and RNA and in the possible control of gene activity.
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PMID:A comparison of the loosely bound and tightly bound nonhistone proteins from Ehrlich ascites tumor chromatin. 97 79


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