Smith Edwin R, Winter Benjamin, Eissenberg Joel C, Shilatifard Ali
Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, MO 64110, USA.
Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8575-9. doi: 10.1073/pnas.0804379105. Epub 2008 Jun 17.
Many developmentally regulated genes contain a poised RNA polymerase II (Pol II) at their promoters under conditions where full-length transcripts are undetectable. It has been proposed that the transcriptional activity of such promoters is regulated at the elongation stage of Pol II transcription. In Drosophila, the heat-shock loci expressing the Hsp70 genes have been used as a model for the regulation of the transcriptional activity of poised Pol II. Drosophila ELL (dELL) is a Pol II elongation factor capable of stimulating the rate of transcription both in vivo and in vitro. Although ELL and the elongation factor Elongin A have indistinguishable effects on RNA polymerase in vitro, the loss-of-function studies indicate that these proteins are not redundant in vivo. In this article, we use RNAi to investigate the physiological properties of dELL and a dELL-associated factor (dEaf) in a living organism. Both ELL and Eaf are essential for fly development. dELL is recruited to heat shock loci upon induction, and its presence with Pol II at such loci is required for proper heat-shock gene expression. Consistent with a role in elongation, dELL knockdown reduces the levels of phosphorylated Pol II at heat-shock loci. This study implicates dELL in the expression of loci regulated by Pol II elongation.
许多受发育调控的基因在无法检测到全长转录本的条件下,其启动子处存在一种就绪的RNA聚合酶II(Pol II)。有人提出,此类启动子的转录活性在Pol II转录的延伸阶段受到调控。在果蝇中,表达热休克蛋白70(Hsp70)基因的热休克基因座已被用作调控就绪Pol II转录活性的模型。果蝇ELL(dELL)是一种Pol II延伸因子,能够在体内和体外刺激转录速率。尽管ELL和延伸因子延伸素A在体外对RNA聚合酶具有难以区分的作用,但功能丧失研究表明,这些蛋白质在体内并非冗余。在本文中,我们使用RNA干扰技术在活体生物中研究dELL和一种与dELL相关的因子(dEaf)的生理特性。ELL和Eaf对果蝇发育均至关重要。诱导后,dELL被招募到热休克基因座,并且在这些基因座上与Pol II共存是热休克基因正常表达所必需的。与在延伸过程中的作用一致,敲低dELL会降低热休克基因座处磷酸化Pol II的水平。这项研究表明dELL参与了受Pol II延伸调控的基因座的表达。