Verbeet M P, Klootwijk J, van Heerikhuizen H, Fontijn R D, Vreugdenhil E, Planta R J
Nucleic Acids Res. 1984 Jan 25;12(2):1137-48. doi: 10.1093/nar/12.2.1137.
To identify DNA elements involved in the initiation of rRNA transcription in yeast we located the start site of the rRNA operon of Kluyveromyces lactis and Hansenula wingei, both members of the Saccharomycetoideae, by S1 nuclease analysis and determined the surrounding nucleotide sequences. Comparison of these sequences with those of Saccharomyces carlsbergensis, S. cerevisiae and S. rosei (all belonging to the same yeast subfamily) reveals an identical sequence at the site of transcription initiation from position +1 to +7 which is part of a larger conserved region extending from position -9 to +23; the conserved heptanucleotide sequence is supposed to constitute an important part of the promoter for yeast RNA polymerase A. The non-transcribed spacers (NTS) upstream of position -9 have diverged strongly with the exception of two short elements around positions -75 and -135. The external transcribed spacer (ETS) downstream of position +23 is largely conserved between K. lactis, S. rosei and S. carlsbergensis except for a divergent region around position +75. On the other hand, the ETS of H. wingei has diverged significantly.
为了鉴定参与酵母中rRNA转录起始的DNA元件,我们通过S1核酸酶分析定位了乳酸克鲁维酵母和汉逊德巴利酵母(均为酵母亚科成员)的rRNA操纵子的起始位点,并确定了其周围的核苷酸序列。将这些序列与卡尔斯伯酵母、酿酒酵母和玫瑰酵母(均属于同一酵母亚科)的序列进行比较,发现在转录起始位点从+1到+7的位置存在相同的序列,该序列是从-9到+23位置延伸的更大保守区域的一部分;保守的七核苷酸序列被认为构成酵母RNA聚合酶A启动子的重要部分。-9位置上游的非转录间隔区(NTS)除了在-75和-135位置附近的两个短元件外,已经发生了很大的分歧。+23位置下游的外部转录间隔区(ETS)在乳酸克鲁维酵母、玫瑰酵母和卡尔斯伯酵母之间基本保守,除了在+75位置附近的一个分歧区域。另一方面,汉逊德巴利酵母的ETS已经有了显著的分歧。