Hansen K, Birse C E, Proudfoot N J
Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, UK.
EMBO J. 1998 Jun 1;17(11):3066-77. doi: 10.1093/emboj/17.11.3066.
We have determined the extent of the primary transcription unit for the two highly expressed genes nmt1 and nmt2 of Schizosaccharomyces pombe. Transcription run-on analysis in permeabilized yeast cells was employed to map polymerase density across the 3'-flanking region of these two genes. Surprisingly, polymerases were detected 4.3 kb beyond the nmt1 polyadenylation [poly(A)] site and 2.4 kb beyond the nmt2 poly(A) site, which in each case have transcribed through an entire convergent downstream transcription unit. However, the steady-state levels of both downstream genes were unaffected by the high level of nmt1 or nmt2 nascent transcription. Analysis of nmt1 and nmt2 RNA 3' end formation signals indicates that efficient termination of transcription requires not only a poly(A) signal but also additional pause elements. The absence of such pause elements close to the poly(A) sites of these genes may account for their extended nascent transcripts.
我们已经确定了粟酒裂殖酵母两个高表达基因nmt1和nmt2的初级转录单元的范围。利用通透化酵母细胞中的转录延伸分析来绘制这两个基因3'侧翼区域的聚合酶密度图。令人惊讶的是,在nmt1聚腺苷酸化[poly(A)]位点下游4.3 kb处以及nmt2 poly(A)位点下游2.4 kb处检测到了聚合酶,在每种情况下,它们都转录通过了一个完整的反向下游转录单元。然而,两个下游基因的稳态水平不受nmt1或nmt2新生转录高水平的影响。对nmt1和nmt2 RNA 3'末端形成信号的分析表明,转录的有效终止不仅需要一个poly(A)信号,还需要额外的暂停元件。这些基因的poly(A)位点附近缺乏此类暂停元件可能解释了它们延长的新生转录本。