SPECIFICITY OF RNA MATURATION PATHWAYS - RNAS TRANSCRIBED BY RNA POLYMERASE-III ARE NOT SUBSTRATES FOR SPLICING OR POLYADENYLATION
SPECIFICITY OF RNA MATURATION PATHWAYS - RNAS TRANSCRIBED BY RNA POLYMERASE-III ARE NOT SUBSTRATES FOR SPLICING OR POLYADENYLATION
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DOI：
10.1128/mcb.7.10.3602
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发表时间：
1987-10-01
影响因子：
5.3
通讯作者：
CLEVELAND, DW
中科院分区：
文献类型：
--
作者：
SISODIA, SS;SOLLNERWEBB, B;CLEVELAND, DW
为了分析RNA加工反应的特异性，我们构建了含有RNA聚合酶III启动子的杂交基因，该启动子与通常由聚合酶II转录的序列融合，并在转染人293细胞后评估其转录物。通过使用组合的RNase H和S1核酸酶测定来分析来自这些嵌合构建体的转录物，以测试含有共有5“和3”剪接信号的RNA是否可以在完整细胞中有效地剪接，即使它们被RNA聚合酶III转录。我们发现聚合酶III衍生的RNA不是剪接的底物。类似地，我们不能检测到来自含有聚合酶III启动子的基因的poly（A）+RNA，该启动子连接到当被RNA聚合酶II转录时指导3“末端切割和聚腺苷酸化所必需和足够的序列。我们的研究结果是一致的，认为在体内剪接和多聚腺苷酸化途径是强制性耦合到RNA聚合酶II的转录。
To analyze the specificity of RNA processing reactions, we constructed hybrid genes containing RNA polymerase III promoters fused to sequences that are normally transcribed by polymerase II and assessed their transcripts following transfection into human 293 cells. Transcripts derived from these chimeric constructs were analyzed by using a combined RNase H and S1 nuclease assay to test whether RNAs containing consensus 5'' and 3'' splicing signals could be efficiently spliced in intact cells, even though they were transcribed by RNA polymerase III. We found that polymerase III-derived RNAs are not substrates for splicing. Similarly, we were not able to detect poly(A)+ RNAs derived from genes that contained a polymerase III promoter linked to sequences that were necessary and sufficient to direct 3''-end cleavage and polyadenylation when transcribed by RNA polymerase II. Our findings are consistent with the view that in vivo splicing and polyadenylation pathways are obligatorily coupled to transcription by RNA polymerase II.