PRODUCTION OF MESSENGER-RNA IN CHINESE-HAMSTER CELLS - RELATIONSHIP OF THE RATE OF SYNTHESIS TO THE CYTOPLASMIC CONCENTRATION OF 9 SPECIFIC MESSENGER-RNA SEQUENCES
PRODUCTION OF MESSENGER-RNA IN CHINESE-HAMSTER CELLS - RELATIONSHIP OF THE RATE OF SYNTHESIS TO THE CYTOPLASMIC CONCENTRATION OF 9 SPECIFIC MESSENGER-RNA SEQUENCES
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DOI：
10.1016/0092-8674(79)90341-6
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发表时间：
1979-01-01
期刊：
影响因子：
64.5
通讯作者：
DARNELL, JE
中科院分区：
文献类型：
--
作者：
HARPOLD, MM;EVANS, RM;DARNELL, JE
从中国仓鼠卵巢[CHO]细胞中构建了与mRNA互补的克隆DNA序列[poly（A）+胞质RNA]，并将其用于RNA：DNA杂交实验。每个克隆的DNA与长度为1.3-3.5 kb的单个mRNA杂交。与各DNA片段互补的核RNA的相对标记率（转录率）不同。十倍在细胞平衡标记后，mRNA中相同特异性RNA序列的相对细胞质浓度发生变化。一百倍还估计了这些细胞质mRNA的核RNA前体分子的大小。一级RNA转录物大小为2.4- 13.5kb，比mRNA大2-6倍，每个克隆的DNA片段只与一种mRNA互补;对于与9个克隆的DNA片段中的至少3个互补的RNA，相对细胞质含量与核RNA合成的相对速率显著不同，表明转录后事件参与了某些哺乳动物mRNA胞质浓度的决定;与9个克隆的DNA片段互补的总非核糖体核RNA的分数在大多数情况下为10- 10%。倍小于细胞质mRNA的部分互补相同的克隆的DNA片段，这表明许多HN[异源核]RNA分子的合成是定性不同于那些最终贡献mRNA的细胞质。
Cloned DNA sequences complementary to unselected mRNA [poly(A)+ cytoplasmic RNA] were constructed from Chinese hamster ovary [CHO] cells and used them in RNA:DNA hybridization experiments. Each cloned DNA hybridized a single mRNA from 1.3-3.5 kb [kilobase] in length. The relative rates of labeling (transcription rates) of nuclear RNA complementary to each individual DNA segment varied .apprx. 10-fold. The relative cytoplasmic concentration of the same specific RNA sequences in the mRNA after an equilibrium labeling of the cells varied .apprx. 100-fold. The sizes of the nuclear RNA precursor molecules to these cytoplasmic mRNA were also estimated. The primary RNA transcripts, which range in size from 2.4-13.5 kb, are 2-6 times larger than the mRNA; each cloned DNA segment is complementary to only 1 species of mRNA; for the RNA complementary to at least 3 of the 9 cloned DNA segments, the relative cytoplasmic content is considerably different from the relative rate of nuclear RNA synthesis, suggesting that post-transcriptional events are involved in the determination of the cytoplasmic concentrations of some mammalian mRNA; and the fraction of total nonribosomal nuclear RNA complementary to 9 cloned DNA segments is in most cases 10-fold less than the fraction of cytoplasmic mRNA complementary to the same cloned DNA segments, suggesting the synthesis of many hn[heterogenous nuclear]RNA molecules that are qualitatively different from those which eventually contribute mRNA to the cytoplasm.