RNA synthesis in isolated nuclei of Xenopus laevis emproyos.
Nuclei were isolated from tailbud embryos of Xenopus laevis and their RNA synthetic activity was studied. The activity was sensitive to actinomycin D, and required the presence of all four ribonucleotide triphosphates and a divalent cation (Mg2+ or Mn2+). The effect of the divalent cation varied depending on the presence of 0.4 M KCl: in its absence more RNA was synthesized with Mg2+ than with Mn2+, but in its presence the situation was reversed. About 50% of the RNA synthesized was sensitive to the presence of alpha-amanitin. Polyacrylamide gel electrophoresis of the labeled RNA revealed that the RNA synthesized had a very heterogeneous size distribution ranging from 40 to 4 S, with a broad peak at around 28--18 S. The RNA which was distributed at around 28--18 S and 4 S was resistant to the presence of a low dose of alpha-amanitin, suggesting its ribosomal and soluble RNA nature. The RNA which was sensitive to this drug and hence may be assumed to be messenger RNA was distributed mainly between 18 and 4 S. From these results it appears that the isolated embryonic nuclei retain their activity to synthesize all of the major classes of RNA.
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D003609:Dactinomycin / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D012313:RNA / Q000096:biosynthesis*; D012335:RNA, Ribosomal / Q000096:biosynthesis; D012343:RNA, Transfer / Q000096:biosynthesis; D014981:Xenopus
D000546:Amanitins; D012335:RNA, Ribosomal; D003609:Dactinomycin; D008345:Manganese; D012313:RNA; D011189:Potassium Chloride; D012343:RNA, Transfer; D008274:Magnesium