Giuffrida A M, Cambria A, Serra I, Avitabile M
Ital J Biochem. 1975 Sep-Oct;24(5):288-307.
RNA polymerase activities of whole nuclei, of isolated and purified nucleoli and of the nucleoplasmic fractions obtained from cerebral hemispheres, cerebellum and brain stem of rat at different days of postnatal development have been determined. In the whole nuclei the fraction of RNA polymerase which is sensitive to alpha-amanitin, is strongly affected by salt concentration; at low ionic strength most of the activity is resistant to the drug while at high ionic strength the enzymatic activity shows a greater sensitivity to the drug. In isolated nucleoli RNA synthesis is not inhibited at all by alpha-amanitin. The biosynthesis of RNA, at low ionic strength, is inhibited by low doses of actinomycin D, whereas at high ionic strength it is remarkably inhibited only by higher doses of the drug. The sensitivity of the reaction to alpha-amanitin and actinomycin D provide good evidence that UTP or GTP incorporation into RNA in purified nuclei and nucleoli, is dependent on RNA polymerases acting on DNA template and is not dependent on homopolymer formation. These results show that in the whole brain nuclei at low ionic strength there is a preferential synthesis of rRNA, whereas at high ionic strength the synthesis of heterogenous RNA predominates. In isolated nucleoli the synthesis of RNA is restricted to rRNA.
已测定了出生后不同发育阶段大鼠大脑半球、小脑和脑干的全细胞核、分离纯化的核仁以及核质部分的RNA聚合酶活性。在全细胞核中,对α-鹅膏蕈碱敏感的RNA聚合酶部分受盐浓度的强烈影响;在低离子强度下,大部分活性对该药物有抗性,而在高离子强度下,酶活性对该药物表现出更大的敏感性。在分离的核仁中,RNA合成完全不受α-鹅膏蕈碱的抑制。在低离子强度下,RNA的生物合成受到低剂量放线菌素D的抑制,而在高离子强度下,仅在较高剂量的该药物作用下才会受到显著抑制。该反应对α-鹅膏蕈碱和放线菌素D的敏感性充分证明,在纯化的细胞核和核仁中,UTP或GTP掺入RNA依赖于作用于DNA模板的RNA聚合酶,而不依赖于均聚物的形成。这些结果表明,在低离子强度下,全脑细胞核中rRNA的合成占优势,而在高离子强度下,异源RNA的合成占主导。在分离的核仁中,RNA的合成仅限于rRNA。