Labhart P
Scripps Research Institute, Department of Molecular and Experimental Medicine, La Jolla, CA 92037, USA.
Nucleic Acids Res. 1995 Jun 25;23(12):2252-8. doi: 10.1093/nar/23.12.2252.
In Xenopus, termination by RNA polymerase I (pol I) is mediated by the 9 bp sequence GACTTGCNC and RNA 3'-ends are formed -15-20 nt upstream of this terminator element. Here I show that this 9 bp element, also called the 'T3 box', is a pause signal for the elongating transcription complex. The two major transcripts in the paused complex have 3'-ends mapping to 15 and 21 nt upstream of the T3 box, remain bound to the template in 0.25% Sarkosyl, are subject to pyrophosphorolysis and can be chased into longer transcripts. Mutations that reduce overall termination also affect pausing, indicating that pausing is a limiting step in the termination process. Oligonucleotide competition experiments, furthermore, suggest that pausing requires a DNA binding factor. The data support a model in which the first step leading to transcription termination by pol I in Xenopus is pausing of the elongation complex upstream of the T3 box.
在非洲爪蟾中,RNA聚合酶I(pol I)介导的转录终止由9个碱基对的序列GACTTGCNC介导,RNA 3'端在该终止元件上游-15-20个核苷酸处形成。在此,我表明这个9个碱基对的元件,也称为“T3框”,是延伸转录复合物的一个暂停信号。暂停复合物中的两个主要转录本的3'端定位于T3框上游15和21个核苷酸处,在0.25%的十二烷基肌氨酸钠中仍与模板结合,易发生焦磷酸解,并且可以被追踪为更长的转录本。降低总体终止的突变也会影响暂停,这表明暂停是终止过程中的一个限制步骤。此外,寡核苷酸竞争实验表明,暂停需要一种DNA结合因子。这些数据支持了一个模型,即非洲爪蟾中pol I导致转录终止的第一步是延伸复合物在T3框上游暂停。