Nascent-seq indicates widespread cotranscriptional pre-mRNA splicing in Drosophila
Nascent-seq indicates widespread cotranscriptional pre-mRNA splicing in Drosophila
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DOI：
10.1101/gad.178962.111
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发表时间：
2011-12-01
影响因子：
10.5
通讯作者：
Rosbash, Michael
中科院分区：
文献类型：
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作者：
Khodor, Yevgenia L.;Rodriguez, Joseph;Rosbash, Michael
为了确定在果蝇中共转录剪接的流行，我们测序了果蝇S2细胞以及果蝇头部的新生RNA转录本。87%的内含子分析表明>50%的共转录剪接。剩下的13%是共转录剪接差或缓慢，与类似的3%几乎完全保留在新生的前mRNA。虽然个别内含子显示轻微的，但统计学上显着的差异剪接效率，类似的全球剪接水平从两个来源。重要的是，具有低共转录剪接效率的内含子存在于具有有效剪接的内含子的相同初级转录物中，表明剪接是内含子特异性的。分析还表明，共转录剪接是效率较低的第一内含子，较长的内含子，和内含子注释为替代。最后，S2细胞表达的慢RpII215（C4）突变体显着减少内含子保留比野生型S2细胞。
To determine the prevalence of cotranscriptional splicing in Drosophila, we sequenced nascent RNA transcripts from Drosophila S2 cells as well as from Drosophila heads. Eighty-seven percent of the introns assayed manifest >50% cotranscriptional splicing. The remaining 13% are cotranscriptionally spliced poorly or slowly, with similar to 3% being almost completely retained in nascent pre-mRNA. Although individual introns showed slight but statistically significant differences in splicing efficiency, similar global levels of splicing were seen from both sources. Importantly, introns with low cotranscriptional splicing efficiencies are present in the same primary transcript with efficiently spliced introns, indicating that splicing is intron-specific. The analysis also indicates that cotranscriptional splicing is less efficient for first introns, longer introns, and introns annotated as alternative. Finally, S2 cells expressing the slow RpII215(C4) mutant show substantially less intron retention than wild-type S2 cells.