DNA-dependent RNA polymerase II from nuclei of suspension-cultured tobacco cells |
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Authors: | Jan Szopa Karl G Wagner |
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Institution: | (1) Abtellung Molekularbiologie, Gesellschaft für Biotechnologische Forschung, Mascheroder Weg 1, D-3300 Braunschweig, Germany |
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Abstract: | From isolated nuclei of suspension cultured cells of Nicotiana tabacum. DNA-dependent RNA polymerase II (E.C. 2.7.76) has been purified to homogeneity as evidenced by polyacrylamidegel electrophoresis under non-denaturing conditions. The purified enzyme had a specific activity of more than 15 nmol min-1·mg-1 with denatured calf thymus DNA as template. Sodium-dodecyl-sulfate gel electrophoresis and protein highperformance liquid chromatography revealed a subunit composition of four proteins with molecular weights of 165 000, 135 000, 35 000 and 25 000 and with a stoichiometry of 1:1:2:2. The RNA polymerase did not exhibit any detectable proteinkinase activity. The 25 000 subunit binds ADP in a molar ratio of 1:1; it could not be decided whether this subunit has an ATPase activity or is merely an acceptor of ADP.Abbreviations HPLC
high-performance liquid chromatography
- PMSF
phenylmethylsulfonyl fluoride
- SDS
sodium dodecyl sulfate
This contribution is dedicated to Professor Fritz Cramer on the occasion of his 60th birthday |
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Keywords: | Nicotiana (RNA polymerase II) RNA polymerase II |
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