PMID: 1191701Nov 18, 1975Paper

DNA-directed RNA polymerase from HeLa cells. Isolation, characterization and cell-cycle distribution of three enzymes.

Biochimica Et Biophysica Acta
B J Benecke, K H Seifart

Abstract

DNA-directed RNA polymerase was solubilized from total HeLa cells. Three distinct classes of the enzyme could be clearly differentiated by their sensitivity toward alpha-amanitin. While form A is completely resistant to high concentrations (133 mug/ml) of this toxin, enzyme B is highly sensitive and is completely inhibited by concentrations of 0.1 mug/ml. In contrast, RNA polymerase C shows an intermediate behaviour (50% inhibition at 30% mug/ml). Separation of the three individual enzymes was achieved by chromatography on DEAE-cellulose (to separate enzyme B from A and C) and DEAE-Sephadex (to separate polymerase A from C). All three RNA polymerases were subsequently purified by phosphocellulose chromatography followed by sedimentation through glycerol gradients. Analysis of the purified enzymes by gel electrophoresis under denaturating conditions showed that the A enzyme consists of five subunits with molecular weights of 185, 128, 65, 41 and 32 X 10(3). In contrast, polymerase B is composed of seven subunits in variable stoichiometry with molecular weights of 215, 175, 145, 123, 68, 43 and 31 X 10(3) respectively. The subunit structure of enzyme C is not entirely clear at present and remains to be established. In addition, R...Continue Reading

References

Jan 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·V E SklarR G Roeder
May 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·R Weinmann, R G Roeder
Mar 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·J SebastianH O Halvorson
Mar 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·R G Roeder, W J Rutter
Apr 3, 1970·Science·H Fan, S Penman
May 28, 1971·Science·E A Zylber, S Penman

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Citations

Jan 1, 1978·The International Journal of Biochemistry·C A Maniglia, R G Wilson
Jan 1, 1982·The International Journal of Biochemistry·R VoetsW Dierick
Jan 1, 1985·CRC Critical Reviews in Biochemistry·A Sentenac
Mar 1, 1977·European Journal of Biochemistry·F Bieri-Bonniot, C Dierks-Ventling

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