Parikh V S, Conrad-Webb H, Docherty R, Butow R A
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235.
Mol Cell Biol. 1989 May;9(5):1897-907. doi: 10.1128/mcb.9.5.1897-1907.1989.
We have identified stable transcripts from the so-called nontranscribed spacer region (NTS) of the nuclear ribosomal DNA repeat in certain respiration-deficient strains of Saccharomyces cerevisiae. These RNAs, which are transcribed from the same strand as is the 37S rRNA precursor, are 500 to 800 nucleotides long and extend from the 5' end of the 5S rRNA gene to three major termination sites about 1,780, 1,830, and 1,870 nucleotides from the 3' end of the 26S rRNA gene. A survey of various wild-type and respiration-deficient strains showed that NTS transcript abundance depended on the mitochondrial genotype and a single codominant nuclear locus. In strains with that nuclear determinant, NTS transcripts were barely detected in [rho+] cells, were slightly more abundant in various mit- derivatives, and were most abundant in petites. However, in one petite that was hypersuppressive and contained a putative origin of replication (ori5) within its 757-base-pair mitochondrial genome, NTS transcripts were no more abundant than in [rho+] cells. The property of low NTS transcript abundance in the hypersuppressive petite was unstable, and spontaneous segregants that contained NTS transcripts as abundant as in the other petites examined could be obtained. Thus, respiration deficiency per se is not the major factor contributing to the accumulation of these unusual RNAs. Unlike RNA polymerase I transcripts, the abundant NTS RNAs were glucose repressible, fractionated as poly(A)+ RNAs, and were sensitive to inhibition by 10 micrograms of alpha-amanitin per ml, a concentration that had no effect on rRNA synthesis. Abundant NTS RNAs are therefore most likely derived by polymerase II transcription.
我们在酿酒酵母的某些呼吸缺陷型菌株中,鉴定出了来自核糖体DNA重复序列所谓非转录间隔区(NTS)的稳定转录本。这些RNA与37S rRNA前体转录自同一条链,长度为500至800个核苷酸,从5S rRNA基因的5'端延伸至距26S rRNA基因3'端约1,780、1,830和1,870个核苷酸处的三个主要终止位点。对各种野生型和呼吸缺陷型菌株的调查表明,NTS转录本丰度取决于线粒体基因型和一个单一的共显性核基因座。在具有该核决定因素的菌株中,在[rho +]细胞中几乎检测不到NTS转录本,在各种mit-衍生物中略为丰富,而在小菌落中最为丰富。然而,在一个超抑制性小菌落中,其757个碱基对的线粒体基因组中含有一个假定的复制起点(ori5),NTS转录本并不比[rho +]细胞中丰富。超抑制性小菌落中NTS转录本丰度低的特性不稳定,可以获得与其他检测的小菌落中一样丰富的NTS转录本的自发分离株。因此,呼吸缺陷本身并不是导致这些异常RNA积累的主要因素。与RNA聚合酶I转录本不同,丰富的NTS RNA可被葡萄糖抑制,分级分离为聚(A)+ RNA,并且对每毫升10微克的α-鹅膏蕈碱抑制敏感,该浓度对rRNA合成没有影响。因此,丰富的NTS RNA很可能是由聚合酶II转录产生的。