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Mechanisms of Antisense Transcription Initiation from the 3′ End of the GAL10 Coding Sequence In Vivo

机译:GAL10编码序列的3'端体内反义转录起始的机制

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In spite of the important regulatory functions of antisense transcripts in gene expression, it remains unknown how antisense transcription is initiated. Recent studies implicated RNA polymerase II in initiation of antisense transcription. However, how RNA polymerase II is targeted to initiate antisense transcription has not been elucidated. Here, we have analyzed the association of RNA polymerase II with the antisense initiation site at the 3′ end of the GAL10 coding sequence in dextrose-containing growth medium that induces antisense transcription. We find that RNA polymerase II is targeted to the antisense initiation site at GAL10 by Reb1p activator as well as general transcription factors (e.g., TFIID, TFIIB, and Mediator) for antisense transcription initiation. Intriguingly, while GAL10 antisense transcription is dependent on TFIID, its sense transcription does not require TFIID. Further, the Gal4p activator that promotes GAL10 sense transcription is dispensable for antisense transcription. Moreover, the proteasome that facilitates GAL10 sense transcription does not control its antisense transcription. Taken together, our results reveal that GAL10 sense and antisense transcriptions are regulated differently and shed much light on the mechanisms of antisense transcription initiation.
机译:尽管反义转录物在基因表达中具有重要的调控功能,但如何启动反义转录仍是未知的。最近的研究暗示RNA聚合酶II参与反义转录的启动。然而,尚未阐明RNA聚合酶II如何靶向起始反义转录。在这里,我们已经分析了RNA聚合酶II与反义起始位点在 GAL10 编码序列的3'端在含葡萄糖的生长培养基中的诱导反义转录的关联。我们发现,RNA聚合酶II通过Reb1p激活剂和 GAL10 的反义起始位点以及用于反义转录起始的一般转录因子(例如TFIID,TFIIB和Mediator)定位。有趣的是,尽管 GAL10 反义转录依赖于TFIID,但其有义转录却不需要TFIID。此外,促进 GAL10 有义转录的Gal4p激活剂对于反义转录是必不可少的。而且,促进 GAL10 有义转录的蛋白酶体不能控制其反义转录。两者合计,我们的结果表明, GAL10 正义和反义转录受到不同的调控,并为反义转录起始机制提供了许多启示。

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