Purification of sea-urchin RNA polymerase II. Characterization by template requirements and sensitivity to inhibitors

The purification of RNA polymerase II from gastrulae of Paracentrotus lividus is described. The enzyme obtained is homogeneous as judged by electrophoresis under non-denaturing conditions. It is able to transcribe both native high-Mr P. lividus DNA and Psammechinus miliaris h22 histone DNA, although single-stranded and nicked DNAs are better templates. P. lividus RNA polymerase II forms with homologous native DNA stable binary complexes that are able to initiate RNA chains after exposure to heparin. Heparin-resistant complexes do not form on nicks of DNA molecules. Sensitivity of sea-urchin RNA polymerase II to rifamycin derivatives and alpha-amanitin has been determined.