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

---

Query: EC:2.7.7.6 (RNA polymerase)
34,946 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Kinetics of RNA chain elongation catalyzed by wheat germ RNA polymerase II have been studied using various synthetic DNA templates in the presence of excess dinucleotide monophosphate primers. With single- or double-stranded homopolymer templates, the double reciprocal plots 1/(velocity) as a function of 1/(nucleotide substrate) exhibit positive, negative or no curvature. With poly(dAT) as template, the mechanism of nucleoside monophosphate incorporation into RNA is not the ping-pong kinetic mechanism which was derived for E. coli RNA polymerase (6). Noncomplementary nucleoside triphosphates inhibit RNA transcription allosterically. Cordycepin triphosphate behaves as ATP, and not only inhibits AMP incorporation but also that of UMP and GMP on appropriate templates. The reason for this complex kinetic behavior is not yet understood. Possibilities are raised that there are several nucleoside triphosphate binding sites on wheat germ RNA polymerase II, that additional nucleoside triphosphate dependent enzymatic activities are required for reaction to occur or that the Km value for incorporation of a given nucleoside monophosphate into RNA is dependent on the length of the RNA chain and/or the nucleotide sequence surrounding the complementary base on the DNA template.
...
PMID:Complex RNA chain elongation kinetics by wheat germ RNA polymerase II. 620 28

The Pb2+ and Zn2+ ions are efficient catalysts for the polycytidylic acid-directed polymerization of an activated guanylic acid derivative, guanosine 5'-phosphorimidazolide. The products include oligomers of 30 to 40 units in length. The nucleotide residues are predominantly 2'-5' linked when Pb2+ is the catalyst, and predominantly 3'-5' linked in the presence of Zn2+. The significance of these results in the context of the prebiotic evolution of RNA polymerase is discussed.
...
PMID:Efficient metal-ion catalyzed template-directed oligonucleotide synthesis. 624 62

It was found that the previously established inhibition of DNA transcription in vitro caused by aminomethylol compounds depends on the suppression of the RNA chains initiation beginning with adenylic but not with guanylic acid. The RNA polymerase binding to DNA is not impaired thereby; the enzyme affinity for the binding sites and RNA elongation also remain unaffected. A comparison of aminomethylol compounds with other alkylating agents demonstrated that the former are the only known alkylating compounds which specifically influence the initiation step. The nature of DNA structural changes (initiation inhibition) caused by aminomethylol compounds is discussed.
...
PMID:[Aminomethylol compounds inhibit the initiation of RNA chain synthesis beginning with adenylic nucleotides]. 631 77

mRNA guanylyltransferase has been extensively purified from calf thymus. A GTP-binding assay was used based on the observations by Shuman and Hurwitz (1981) and Venkatesan and Moss (1982) that vaccinia virus and HeLa cell mRNA guanylyltransferases bind the GMP moiety from GTP in the absence of an acceptor RNA. The mol. wt. of the purified enzyme from calf thymus, estimated by polyacrylamide gel electrophoresis in the presence of SDS, is 65 000. The major protein in the purified enzyme fraction comigrates with the peptide labelled with GMP. Based on scans of silver-stained polyacrylamide gels, mRNA guanylyltransferase constitutes greater than 50% of the protein in these fractions. The enzyme catalyzed the guanylylation at the 5' end of poly(A) with a mixture of diphosphate and triphosphate ends. No evidence was obtained for a direct interaction between mRNA guanylyltransferase and RNA polymerase B (II).
...
PMID:Purification of mRNA guanylyltransferase from calf thymus. 632 86

The heterogeneity of cell size of E. coli WU-36-10-11-12 and its four RNA-polymerase (rif-r) mutants with pleiotropic effect -- rpoB401, rpoB402, rpoB403 and rpoB409 was investigated for the purposeful choice of E. coli mutant with an altered fidelity of transcription. The stability of the phenotype of E. coli strains was shown to depend on the structural state of RNA polymerase. In vitro RNA-polymerase of the morphologically most unstable mutant rpoB402 incorporates non-complementary GMP or CMP on the poly [d(AT).d(AT)] template more frequently than the enzyme from the wild-type strain. The data obtained suggest that the beta-subunit of RNA-polymerase determines the fidelity of transcription and the selection of complementary nucleotides.
...
PMID:[Role of RNA polymerase in ensuring fidelity in copying the template during transcription in E. coli]. 702 23

The ATP analog 5'-adenylyl imidodiphosphate (AMP-PNP) inhibits transcription of specific genes by the RNA polymerase II contained in whole cell extracts, not only with promoters that contain A as the first nucleotide of the transcript, but also with those that initiate transcripts with G or U. The analog AMP-PNP (a competitive inhibitor of ATP) probably acts at the level of initiation of transcription, but it can be used for elongation by RNA polymerase II in isolated nuclei or in the whole cell extract. AMP-PNP and the other imidotriphosphates have little effect on purified HeLa cell RNA polymerase II initiation and elongation of transcription. Since RNA polymerase III in the crude system both initiates and elongates transcripts with AMP-PNP, we conclude that the availability of the beta-gamma bond of ATP is an indispensable requirement for faithful and specific in vitro initiation only by RNA polymerase II in the whole cell extract. Uncapped U- or G-initiated transcripts were obtained in the presence of UMP-PNP or GMP-PNP, the respective imidodiphosphate analogs. The presence of the 5'-terminal imidotriphosphate at the same oligonucleotide as the cap for U-initiated precursors established that transcription initiation and capping occur at the same site. Capping is not required for transcription by RNA polymerase II in the in vitro system. Methylation of the 2' ribose of the initiating nucleotide does not occur on the imidonucleotide containing 5' ends of adenovirus EIV or murine leukemia virus long terminal repeat.
...
PMID:Mechanism of RNA polymerase II--specific initiation of transcription in vitro: ATP requirement and uncapped runoff transcripts. 715 Nov 73

Transcription termination by vaccinia virus RNA polymerase during synthesis of early mRNAs requires a virus-encoded termination factor (VTF). VTF is but one of many activities associated with the vaccinia virus mRNA capping enzyme, a heterodimer of 95- and 33-kDa subunits encoded by the D1 and D12 genes, respectively. Although the three catalytic domains involved in cap formation have been assigned to individual subunits or portions thereof, the structural requirements for VTF activity are unknown. We now report that both full-length subunits are required for transcription termination. The 844-amino acid D1 subunit by itself, which is fully active in triphosphatase and guanylyltransferase functions, has no demonstrable VTF activity in vitro. Neither does the D12 subunit by itself. The heterodimeric methyltransferase domain of D1 (residues 498 to 844) and D12 subunits also has no VTF activity. VTF is not affected by a K-to-M mutation of the guanylyltransferase active site at position 260 (K260M) that abolishes enzyme-GMP complex formation or by a H682A/Y683A double mutation of the D1 subunit, which abrogates methyltransferase activity. Thus, the structural requirements for termination are distinct from those for nucleotidyl transfer and methyl transfer.
...
PMID:The D1 and D12 subunits are both essential for the transcription termination factor activity of vaccinia virus capping enzyme. 774 34

Avian reovirus S1133 was shown to contain all the enzymatic activities required for the synthesis of mature viral transcripts, including a dsRNA-dependent RNA polymerase, a nucleoside triphosphate phosphohydrolase, an mRNA guanylyltransferase, and two mRNA methyltransferases. The virus used these enzymes both in vitro and in vivo to catalyze the synthesis of viral mRNAs containing a type-1 cap at their 5' ends. Incubation of reovirions with GTP led to the formation of an intermediate structure consisting of GMP bound to the viral core protein lambda 3 through a phosphoamide linkage. The reaction was specific for GTP and required the presence of both Mg2+ and inorganic pyrophosphatase. The GMP moiety can be transferred from the lambda 3-GMP complex to acceptors such as GDP and GTP, yielding GpppG and GppppG, respectively. Our results demonstrate that lambda 3 is the avian reovirus guanylyltransferase.
...
PMID:Endogenous enzymatic activities of the avian reovirus S1133: identification of the viral capping enzyme. 785 76

The double-stranded RNA bacteriophage phi 6 contains a virion-associated RNA-dependent RNA polymerase complex. Removal of the virus envelope and the nucleocapsid surface protein, P8, reveals a nucleocapsid core particle (proteins P1, P2, P4, P7) which is the viral polymerase complex, capable of synthesizing RNA strands of positive polarity. The in vitro plus strand synthesis (transcription) reaction of the particle obtained from the mature virion was optimized and its activation and inactivation were investigated. Purine nucleoside triphosphates (NTPs), binding to a low-affinity binding site in the polymerase complex, activated plus strand synthesis. GTP was the preferred NTP, but dGTP, ddGTP, and the noncleavable analog GMP-PCP could also switch on transcription. This NTP-binding site is probably different from that of the unspecific viral NTPase found in protein P4 and also from that of the rNTP-specific RNA polymerase active site. Binding of purine NTPs was sufficient for the switch-on; hydrolysis of the NTP was not required. Besides nucleotides, divalent cations had an effect on phi 6 in vitro plus strand synthesis. Magnesium ions are required for the activity but calcium ions inhibit the reaction. Manganese ions are shown to dissipate the effect of magnesium and calcium ions, leading to uncontrolled, exceptionally high level plus strand synthesis.
...
PMID:In vitro transcription of the double-stranded RNA bacteriophage phi 6 is influenced by purine NTPs and calcium. 788 44

The Escherichia coli GreA and GreB proteins induce cleavage of 3' fragments from nascent transcripts in halted transcription complexes. We have overproduced and purified the GreA protein and tested how it affects initiation, pausing, and termination by E. coli RNA polymerase. Recombinant GreA induced cleavage of two to three nucleotide fragments in two promoter-proximal complexes, whereas an apparently endogenous cleavage removed a single larger fragment. Both types of cleavage stopped once the transcript was shortened to approximately 10 nucleotides. However, during initiation, GreA induced cleavage of transcripts as short as four nucleotides, inhibiting their release as abortive products and stimulating both productive initiation and "primer-shifting" at a weak promoter. GreA induced repetitive cleavage over a long distance in complexes containing a long G-less nascent transcript. However, reverse translocation was inhibited in transcription complexes that contained a G-rich, C-less nascent transcript. Substituting IMP for GMP in the transcript relieved inhibition. Finally, GreA had little effect on transcription through the his and trp leader pause sites or on termination at nine different p-independent terminators. We propose that transcript cleavage and reverse translocation are controlled in part by backsliding of the nascent transcript through an RNA-binding site.
...
PMID:GreA-induced transcript cleavage in transcription complexes containing Escherichia coli RNA polymerase is controlled by multiple factors, including nascent transcript location and structure. 807 55


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---