Kawauchi Junya, Mischo Hannah, Braglia Priscilla, Rondon Ana, Proudfoot Nick J
Sir William Dunn School of Pathology, Oxford OX1 3RE, United Kingdom.
Genes Dev. 2008 Apr 15;22(8):1082-92. doi: 10.1101/gad.463408.
Both RNA polymerase I and II (Pol I and Pol II) in budding yeast employ a functionally homologous "torpedo-like" mechanism to promote transcriptional termination. For two well-defined Pol II-transcribed genes, CYC1 and PMA1, we demonstrate that both Rat1p exonuclease and Sen1p helicase are required for efficient termination by promoting degradation of the nascent transcript associated with Pol II, following mRNA 3' end processing. Similarly, Pol I termination relies on prior Rnt1p cleavage at the 3' end of the pre-rRNA 35S transcript. This is followed by the combined actions of Rat1p and Sen1p to degrade the Pol I-associated nascent transcript that consequently promote termination in the downstream rDNA spacer sequence. Our data suggest that the previously defined in vitro Pol I termination mechanism involving the action of the Reb1p DNA-binding factor to "road-block" Pol I transcription close to the termination region may have overlooked more complex in vivo molecular processes.
出芽酵母中的RNA聚合酶I和II(Pol I和Pol II)都采用功能同源的“鱼雷样”机制来促进转录终止。对于两个明确的Pol II转录基因CYC1和PMA1,我们证明,在mRNA 3'末端加工后,Rat1p核酸外切酶和Sen1p解旋酶通过促进与Pol II相关的新生转录本的降解,对于有效终止都是必需的。同样,Pol I终止依赖于Rnt1p在pre-rRNA 35S转录本3'末端的先前切割。随后是Rat1p和Sen1p的联合作用,以降解与Pol I相关的新生转录本,从而促进下游rDNA间隔序列中的终止。我们的数据表明,先前定义的涉及Reb1p DNA结合因子作用以在终止区域附近“阻碍”Pol I转录的体外Pol I终止机制可能忽略了更复杂的体内分子过程。