Yu K, Elder R T
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
Mol Cell Biol. 1989 Jun;9(6):2431-44. doi: 10.1128/mcb.9.6.2431-2444.1989.
Fragments from the Ty-D15 element of Saccharomyces cerevisiae were assayed for the ability to direct 3'-end formation for RNA initiated by the GAL1 promoter. The delta, the direct repeat at each end of the element, was capable of forming 3' ends at two sites, an inefficient upstream site and an efficient downstream site near the end of the delta. Different sequences were required for 3'-end formation at these sites. For the efficient site, all transcripts had 3' ends in the delta and no downstream transcription was detected, which suggested that these sequences terminate transcription. Surprisingly, the delta region downstream of the initiation site for Ty RNA comprised part of this major site and terminated more than 50% of the transcripts that read into it. Sequences necessary for the efficient site were localized to two small regions. Both regions were upstream of the 3' end and contained similarities to a tripartite consensus sequence that has been proposed as a terminator element. Sequences near the position of the 3' end could also affect termination; a short G + C-rich sequence inserted just downstream changed an efficient terminator to an inefficient one. Initiation in the delta had no effect on the efficiency or positions or termination in that delta. A new initiation site was seen when the same delta terminated transcription, but transcriptional interference did not occur, since the amount of initiation was not decreased.
对酿酒酵母Ty-D15元件的片段进行了检测,以确定其指导由GAL1启动子起始的RNA 3'末端形成的能力。该元件两端的直接重复序列δ能够在两个位点形成3'末端,一个是效率较低的上游位点,另一个是靠近δ末端的效率较高的下游位点。在这些位点形成3'末端需要不同的序列。对于效率较高的位点,所有转录本的3'末端都在δ中,未检测到下游转录,这表明这些序列可终止转录。令人惊讶的是,Ty RNA起始位点下游的δ区域构成了这个主要位点的一部分,并使超过50%转录进入该区域的转录本终止。高效位点所需的序列定位于两个小区域。这两个区域都在3'末端的上游,并且与已被提议作为终止元件的三联体共有序列相似。3'末端位置附近的序列也会影响终止;就在下游插入的一段富含G + C的短序列将一个高效终止子变成了低效终止子。在δ中的起始对该δ中的终止效率、位置均无影响。当相同的δ终止转录时会出现一个新的起始位点,但由于起始量未减少,因此未发生转录干扰。