==================================BSR50==================================
50.  Structure and isolation of eukaryotic RNA polymerase II or
     RNA polymerase B.
1
UI  - 87096341
AU  - Skripal IG ; Weeks JR ; Greenleaf AL
TI  - Dye-ligand affinity chromatography of RNA polymerase II.
AB  - The binding of wheat germ RNA polymerase II to five different dye-ligand
      chromatography gels (Matrex gels, Amicon Corp.) was tested. A
      quantitative binding of the enzyme to four of the gels, namely Dyematrex
      gels Blue A, Blue B, Red A and Green A was observed. Only the Orange A
      gel column failed to bind the enzyme strongly. Nearly 100% of the
      activity could be recovered from the Green A column by elution with high
      salt concentration and high pH. Under these conditions only a part of the
      activity was eluted from the other three columns since the enzyme bound
      tightly. Enzyme activity could be removed from the columns by elution
      with nucleotide substrates, but the yield from the Blue A, Blue B and Red
      A columns was still low (7 to 42%). The Green A Matrex gel appeared to be
      useful for the purification and analysis of RNA polymerase.
MH  - Chromatography, Affinity/*METHODS ; Chromatography, Agarose ; Dyes ;
      Electrophoresis, Polyacrylamide Gel ; RNA Polymerase II/*ISOLATION &
      PURIFICATION ; Support, U.S. Gov't, P.H.S. ; Wheat/ENZYMOLOGY
SO  - Acta Biochim Biophys Hung 1986;21(3):215-24
2
UI  - 87074902
AU  - Garcia-Carranca A ; Miguel F ; Dahmus ME ; Gariglio P
TI  - Structure of monkey kidney cell RNA polymerase II: characterization of
      RNA polymerase associated with SV40 late transcriptional complexes.
AB  - Three subspecies of RNA polymerase II have been described in eucaryotic
      cells and designated IIO, IIA, and IIB. Although their relative
      proportions vary among different sources, RNA polymerases IIA and IIB
      constitute the bulk of most purified RNA polymerase II preparations.
      Antibodies against calf thymus RNA polymerase II were used to estimate
      the amount of polymerase II subspecies in monkey kidney cells, isolated
      nuclei, and SV40 late transcriptional complexes. We have found that RNA
      polymerase IIO is present in whole cells and isolated nuclei in higher
      proportions than previously reported. Subspecies IIO was found associated
      with SV40 minichromosomes engaged in transcription during late lytic
      infection. The observation that RNA polymerase IIO is associated with the
      cellular chromatin and SV40 minichromosomes suggest that this form of the
      enzyme is the subspecies active in in vivo transcription.
MH  - Animal ; Cattle ; Cell Line ; Cell Nucleus/ENZYMOLOGY ; Cercopithecus
      aethiops ; Chromatin/*ENZYMOLOGY ; Chromosomes/ENZYMOLOGY ; Kidney/
      ENZYMOLOGY ; Molecular Weight ; RNA Polymerase II/IMMUNOLOGY/*ISOLATION &
      PURIFICATION ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S.
      ; SV40 Virus ; Thymus Gland/ENZYMOLOGY ; *Transcription, Genetic
SO  - Arch Biochem Biophys 1986 Nov 15;251(1):232-8
3
UI  - 86243294
AU  - Mazus B ; Falchuk KH ; Vallee BL
TI  - Inhibition of Euglena gracilis and wheat germ zinc RNA polymerases II by
      1,10-phenanthroline acting as a chelating agent.
AB  - Copper complexes of 1,10-phenanthroline (OP-Cu) hydrolyze DNA [D'Aurora,
      V., Stern, A. M., & Sigman, D. S. (1978) Biochem. Biophys. Res. Commun.
      80, 1025-1032; Marshall Pope, L., Reich, K. A., Graham, D. R., & Sigman,
      D. S. (1982) J. Biol. Chem. 257, 12121-12128]. This reaction has been
      studied to determine whether the 1,10-phenanthroline (OP) inhibition of
      the activity of RNA and DNA polymerases is the result of template
      hydrolysis or the chelation of a metal associated with and essential to
      the function of these enzymes. Addition of 4',6-diamino-2-phenylindole
      dihydrochloride (DAPI) to DNA generates a fluorescence signal with a
      linear increase of the intensity over a broad range of DNA concentrations
      from 0 to 100 micrograms/mL. The progress of hydrolysis of DNA by DNase I
      or OP (2 mM) is monitored by the time-dependent decrease in DAPI-induced
      fluorescence. In the presence of OP, the rate of hydrolysis increases as
      the Cu2+ concentration in the reaction mixture rises from 10(-8) to
      10(-5) M. The rate differs for each nucleic acid template used;
      hydrolysis of poly(dA-dT) greater than denatured DNA greater than
      double-stranded DNA. However, millimolar amounts of OP do not hydrolyze
      the template even in the presence of Cu2+ (10(-6) M) when DNA is
      complexed with either Escherichia coli DNA polymerase I or Euglena
      gracilis or wheat germ RNA polymerase II. Under the same conditions, OP
      inhibits the activity of both varieties of RNA polymerase II with pKi's
      of 3.4 and 3.0, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - Chelating Agents/*PHARMACOLOGY ; Comparative Study ; Copper/PHARMACOLOGY
      ; Euglena Gracilis/*ENZYMOLOGY ; Kinetics ; Phenanthrolines/*PHARMACOLOGY
      ; Plants/*ENZYMOLOGY ; RNA Polymerase II/*ANTAGONISTS & INHIBITORS ;
      Structure-Activity Relationship ; Substrate Specificity ; Support, U.S.
      Gov't, P.H.S. ; Wheat/ENZYMOLOGY ; Zinc/*ANALYSIS
SO  - Biochemistry 1986 May 20;25(10):2941-5