Groner B, Phillips S L
J Biol Chem. 1975 Jul 25;250(14):5640-6.
A procedure has been designed for the simultaneous isolation, in a single step, of the nuclei and cytoplasm of Saccharomyces cerevisiae alphas288c spheroplasts. We have examined the polyadenylate poly(A)-containing RNA in these fractions and their kinetics of synthesis. Nuclear RNA saturates with [3H] adenine within 10 min. Labeled RNA appears very quickly in the cytoplasm, exceeding the amount of labeled nuclear RNA within 2 min after the addition of [3H] adenine. Nuclear poly(A)-containing RNA is approximately 10% of the total cellular poly(A)-containing RNA obtained from spheroplasts labeled for 15 min. Nuclear poly(A)-containing RNA is not as large as the giant heterogeneous nuclear RNA of animal cells. The distribution of molecular size in nuclear and cytoplasmic populations of poly(A)-containing RNA is very broad with the average size of the nuclear species being moderately larger than the cytoplasmic species. Three distinct size classes of poly(A), with different apparent kinetic properties, are obtained from yeast poly(A)-containing RNA. Their electrophoretic mobility suggests molecular lengths of approximately 20, 40, and 60 nucleotides (Groner, G., Hynes, N., and Phillips, S. (1975) Biochemistry, 13, 5378-5383). Each of these poly(A) classes are present in the mRNA from large and small polyribosomes.
已设计出一种程序，可在单个步骤中同时分离酿酒酵母α288c原生质球的细胞核和细胞质。我们已检测了这些组分中含聚腺苷酸（poly(A)）的RNA及其合成动力学。细胞核RNA在10分钟内用[3H]腺嘌呤饱和。标记的RNA在细胞质中出现得非常快，在加入[3H]腺嘌呤后2分钟内就超过了标记的细胞核RNA的量。从标记15分钟的原生质球获得的细胞核中含poly(A)的RNA约占细胞总含poly(A)RNA的10%。细胞核中含poly(A)的RNA不像动物细胞中的巨大异质核RNA那么大。含poly(A)RNA在细胞核和细胞质群体中的分子大小分布非常广泛，细胞核中的平均大小略大于细胞质中的。从酵母含poly(A)的RNA中获得了三种不同大小类别的poly(A)，它们具有不同的表观动力学特性。它们的电泳迁移率表明分子长度约为20、40和60个核苷酸（格罗纳，G.，海因斯，N.，和菲利普斯，S.（1975年）《生物化学》，13，5378 - 5383）。这些poly(A)类别中的每一种都存在于大小多核糖体的mRNA中。
