Sitzler S, Oldenburg I, Petersen G, Bautz E K
Institut für Molekulare Genetik, Universität Heidelberg, F.R.G.
Gene. 1991 Apr;100:155-62. doi: 10.1016/0378-1119(91)90361-e.
To analyze the transcriptional control regions of Drosophila melanogaster household genes, we have characterized the promoter of the gene coding for the second-largest subunit of RNA polymerase II (DmRP140). Analysis of cDNA revealed that the coding region of the protein extends beyond the originally assumed transcription start point (tsp) and deduced translation start codon [Falkenburg et al., J. Mol. Biol. 195 (1987) 929-937] and that the tsp determined previously corresponds to an intron/exon boundary of an additional intron. Upstream of the polII gene we found a transcription unit that is transcribed in the opposite direction. The initiating ATGs of the two genes are only 467 nucleotides (nt) apart. The untranslated region is extremely A + T-rich (88%) but none of the transcription units is preceded by a canonical TATA element. It does not feature any other known nt sequence motifs thought to be necessary for the basic transcriptional machinery; yet, this region functions as a bidirectional promoter: a central 309-bp fragment directs transcription of a reporter gene in transiently transfected Drosophila culture cells in both orientations. The gene coding for the second-largest subunit of RNA polymerase II of Drosophila virilis (DvRP140) was isolated and partially analyzed. The gene is located on the second chromosome at 22F/23A which corresponds to the position determined for D. melanogaster.(ABSTRACT TRUNCATED AT 250 WORDS)
为了分析黑腹果蝇看家基因的转录控制区域,我们对编码RNA聚合酶II第二大亚基(DmRP140)的基因启动子进行了特征分析。对cDNA的分析表明,该蛋白质的编码区域延伸至最初假定的转录起始点(tsp)和推导的翻译起始密码子之外[Falkenburg等人,《分子生物学杂志》195 (1987) 929 - 937],并且先前确定的tsp对应于一个额外内含子的内含子/外显子边界。在polII基因上游,我们发现了一个以相反方向转录的转录单元。这两个基因的起始ATG仅相隔467个核苷酸(nt)。非翻译区富含A + T(88%),但没有一个转录单元之前有典型的TATA元件。它没有任何其他被认为是基本转录机制所必需的已知nt序列基序;然而,该区域作为双向启动子发挥作用:一个309 bp的中央片段在瞬时转染的果蝇培养细胞中以两个方向指导报告基因的转录。分离并部分分析了果蝇(DvRP140)编码RNA聚合酶II第二大亚基的基因。该基因位于第二条染色体的22F/23A处,这与黑腹果蝇确定的位置相对应。(摘要截短于250字)