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培养的红白血病Friend细胞中珠蛋白mRNA的生物合成与稳定性

Biosynthesis and stability of globin mRNA in cultured erythroleukemic Friend cells.

作者信息

Aviv H, Voloch Z, Bastos R, Levy S

出版信息

Cell. 1976 Aug;8(4):495-503. doi: 10.1016/0092-8674(76)90217-8.

Abstract

Biosynthesis and stability of the mRNA population in DMSO-induced Friend erythroleukemic cells were studied after labeling the RNA with 3H-uridine and then chasing it with nonlabeled uridine. Globin RNA metabolism was studied by hybridization to excess complementary DNA convalently coupled to oligo(dT)-cellulose. After a labeling period of 120 min, 2-4% of the poly(A)-containing labeled RNA was in globin RNA; it decayed with a half-life of 16-17 hr. The rest of the poly(A)-containing RNA was composed to two kinetic populations: 85-90% decayed with a half-life of about 3 hr, while 10% decayed with a half-life of about 37 hr. The portion of globin RNA in labeled poly(A)-containing RNA behaved in an unexpected fashion during the chase period. During the initial chase period, the percentage of globin RNA increased rapidly, reaching a maximum of about 15% at 20 hr, but it subsequently declined gradually. Based on these findings, a model was built that describes the changes in the proportion of globin mRNA in poly(A)-containing RNA during continuous synthesis and after chase of the labeled RNA. It appears that if the parameters described remain constant during the maturation of erythroblasts, then this model would not account for the almost exclusive presence of globin RNA in the reticulocyte. By far the most effective way to achieve this high level of globin RNA is the destabilization of the mRNA population which is more stable than globin RNA, and not the stabilization of globin RNA itself.

摘要

在用³H-尿苷标记RNA后,再用未标记的尿苷进行追踪,研究了二甲基亚砜诱导的Friend红白血病细胞中mRNA群体的生物合成和稳定性。通过与共价偶联到寡聚(dT)-纤维素上的过量互补DNA杂交来研究珠蛋白RNA的代谢。在120分钟的标记期后,含poly(A)的标记RNA中有2-4%是珠蛋白RNA;其半衰期为16-17小时。其余含poly(A)的RNA由两个动力学群体组成:85-90%的半衰期约为3小时,而10%的半衰期约为37小时。在追踪期内,标记的含poly(A)RNA中珠蛋白RNA的部分表现出意想不到的行为。在最初的追踪期内,珠蛋白RNA的百分比迅速增加,在20小时时达到约15%的最大值,但随后逐渐下降。基于这些发现,构建了一个模型,该模型描述了在连续合成和标记RNA追踪后,含poly(A)RNA中珠蛋白mRNA比例的变化。似乎如果在成红细胞成熟过程中所描述的参数保持不变,那么这个模型将无法解释网织红细胞中几乎只存在珠蛋白RNA的现象。到目前为止,实现这种高水平珠蛋白RNA的最有效方法是使比珠蛋白RNA更稳定的mRNA群体不稳定,而不是珠蛋白RNA本身的稳定。

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