Isolation and purification of RNA polymerases from rye embryos

Fabisz-Kijowska, Anna; Dullin, Piotr; Walerych, W.

doi:10.1016/0005-2787(75)90013-1

Cited by 18 publications

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References 26 publications

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“…Plants are known to possess two major forms of RNA polymerase, I and II, and a third form has been described in rye embryos (6). Here, we describe the separation and purification of multiple forms of nuclear RNA polymerases from cauliflower.…”

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Purification and Characterization of DNA-dependent RNA Polymerases from Cauliflower Nuclei

Guilfoyle¹

1976

Plant Physiol.

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DNA-dependent RNA polymerases were solubilized from nuclei of cauliflower inflorescences and purified by agarose A-1.5m, DEAEceflulose, DEAE-Sephadex, and Eukaryotes possess multiple forms of DNA-dependent RNA polymerase which transcribe specific DNA cistrons (3, 4, 10).The major forms of eukaryotic nuclear RNA polymerases can be grouped into three species which are referred to as either I or A, II or B, and III or C (3,4,10). RNA polymerase I is localized within the nucleolus (11,13,18), transcribes ribosomal DNA cistrons (9, 17), and is refractory to the fungal toxin, a-amanitin (11,14). RNA polymerase II is localized within the nucleoplasm (11, 18), synthesizes heterogeneous and presumably messenger RNA (28), and is inhibited by 10-8 to 10-9 Mi a-amanitin (11,14). RNA polymerase III is localized in the nucleoplasm (18,20) and possibly the cytoplasm (2,20,21,24,26), synthesizes ribosomal 5S and transfer RNAs (25), and is inhibited by 10-3 to 10-4 M a-amanitin in higher animals (2,20,21,24,26), but is refractory to the fungal toxin in lower eukaryotes (1,23,27).Most of the studies on eukaryotic RNA polymerases have been conducted with either animals or lower eukaryotes such as yeast, while studies on higher plants have been limited (3). Plants are known to possess two major forms of RNA polymerase, I and II, and a third form has been described in rye embryos (6). Here, we describe the separation and purification of multiple forms of nuclear RNA polymerases from cauliflower. Cauliflower inflorescences offer a rich source of RNA polymerases (7,19). The fractionation of three species of RNA polymerase from cauliflower which have properties similar to RNA polymerases I, II, and III described in animals has been achieved in this work.Although the characteristics of the plant enzymes are in general similar to the animal RNA polymerases, a 10-to 100-fold greater concentration of a-amanitin is required to inhibit cauliflower RNA polymerase III than is required to inhibit the animal form III enzyme (20,23

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