Kempers-Veenstra A E, Oliemans J, Offenberg H, Dekker A F, Piper P W, Planta R J, Klootwijk J
EMBO J. 1986 Oct;5(10):2703-10. doi: 10.1002/j.1460-2075.1986.tb04554.x.
Deletion analysis of artificial rRNA minigenes transformed into Saccharomyces cerevisiae revealed that a 110 bp long fragment corresponding to positions -36 to +74 relative to the 3'-end of the 26S rRNA gene, is both necessary and sufficient for obtaining transcripts whose 3'-termini are identical to those of 26S and 37S (pre-)rRNA. These termini are produced via processing of longer transcripts because in an rna 82.1 mutant the majority of the minigene transcripts extend further downstream. Since the rna 82.1 mutation inactivates an endonuclease involved in the 3'-processing of 5S pre-rRNA it is concluded that the maturation of 37S- and that of 5S pre-rRNA requires a common factor. Comparison of the spacer sequences between Saccharomyces carlsbergensis, Saccharomyces rosei and Hansenula wingei revealed several conserved sequence blocks within the region between +10 and +55. These conserved sequence tracts, which are part of a longer region showing dyad symmetry, are supposed to be involved in the interaction with the processing component(s). Deletion of the sequences required for the formation of the 3'-ends of 26S rRNA and 37S pre-rRNA revealed a putative terminator for transcription by RNA polymerase I situated at position +210. This site maps within a DNA fragment that also contains the enhancing element for rDNA transcription by RNA polymerase I.
对转化到酿酒酵母中的人工rRNA小基因进行缺失分析表明,一个110 bp长的片段,对应于相对于26S rRNA基因3'端的-36至+74位,对于获得其3'末端与26S和37S(前体)rRNA相同的转录本是必要且充分的。这些末端是通过对更长转录本的加工产生的,因为在rna 82.1突变体中,大多数小基因转录本向下游延伸得更远。由于rna 82.1突变使参与5S前体rRNA 3'加工的一种核酸内切酶失活,因此得出结论,37S前体rRNA和5S前体rRNA的成熟需要一个共同因子。对卡尔斯伯酵母、玫瑰酵母和温奇汉逊酵母之间间隔区序列的比较揭示了在+10至+55区域内的几个保守序列块。这些保守序列片段是显示二元对称的更长区域的一部分,被认为参与与加工成分的相互作用。删除26S rRNA和37S前体rRNA 3'末端形成所需的序列,揭示了一个位于+210位的RNA聚合酶I转录的推定终止子。该位点位于一个DNA片段内,该片段还包含RNA聚合酶I对rDNA转录的增强元件。