Purification and subunit structure of RNA polymerase II from different stages of Artemia development

RNA polymerase II has been purified from different stages of Artemia development: cryptobiotic gastrulae, developing embryos and nauplii. RNA polymerase from dormant and developing embryos consists of 12 subunits of 205 000, 140 000, 32 500, 25 600, 22 000, 19 400, 18 000, 16 800, 147 000, 14 000, 12 700 and 11 500 daltons, as determined by dodecyl sulphate polyacrylamide gel electrophoresis. The enzyme from larvae contains the same polypeptides except for the higher-molecular-mass component: the larval enzyme has a subunit of 172 kDa instead of the 205-kDa subunit present in the enzyme purified from embryos. Incubation of embryonic RNA polymerase II with larval extracts results in the conversion of the 205-kDa subunit into the 172-kDa one. This conversion is inhibited by soybean trypsin inhibitor, a compound which inhibits three proteases induced during Artemia early larval development. RNA polymerase II purified from a mixture of embryos and larvae contain only the 172-kDa subunit. Our results indicate that the 172-kDa subunit present in RNA polymerase II purified from larvae is produced by proteolysis of the 205-kDa subunit during the extraction and purification of the enzyme and therefore that Artemia has only one class of RNA polymerase II.