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首页> 外文期刊>Molecular and Cellular Biology >Specificity of RNA maturation pathways: RNAs transcribed by RNA polymerase III are not substrates for splicing or polyadenylation.
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Specificity of RNA maturation pathways: RNAs transcribed by RNA polymerase III are not substrates for splicing or polyadenylation.

机译：RNA成熟途径的特异性：RNA聚合酶III转录的RNA不是剪接或聚腺苷酸化的底物。

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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.
机译：为了分析RNA加工反应的特异性，我们构建了包含RNA聚合酶III启动子的杂合基因，该启动子与通常由聚合酶II转录的序列融合，并在转染到人类293细胞后评估了它们的转录本。通过使用组合的RNase H和S1核酸酶分析法分析了来自这些嵌合构建体的转录本，以测试包含共有5'和3'剪接信号的RNA是否可以在完整细胞中有效剪接，即使它们被RNA聚合酶III转录也是如此。我们发现，聚合酶III衍生的RNA并非用于剪接的底物。同样，我们无法检测到由含有聚合酶III启动子的基因衍生的poly（A）+ RNA，这些启动子与被RNA聚合酶II转录时指导3'末端切割和聚腺苷酸化的序列足够必要。我们的发现与以下观点一致：体内剪接和聚腺苷酸化途径必定与RNA聚合酶II转录结合。

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