Lisowsky T, Michaelis G
Botanisches Institut, Universität Düsseldorf, Federal Republic of Germany.
Mol Gen Genet. 1989 Oct;219(1-2):125-8. doi: 10.1007/BF00261167.
In our previous work (Lisowsky et al. 1987; Lisowsky and Michaelis 1988) we have identified two nuclear pet genes of yeast that are required for mitochondrial transcription. In this report we show that one of these pet mutations, pet-ts798, maps in the RP041 gene encoding mitochondrial RNA polymerase. The temperature-sensitive lesion of mutant pet-ts798 can be suppressed by a second nuclear gene RF1023 (mtf1) when inserted into a high copy number plasmid. Our assumption that mtf1 codes for a 40 kDa mitochondrial transciription factor is supported by the fact that the cloned gene acts as an intergenic suppressor of a temperature-sensitive RNA polymerase mutant. A third nuclear gene (mtf2) for mitochondrial transcription was identified by analysing mutant pet-ts3504. The in vitro transcriptional activity of isolated mutant mitochondria is temperature sensitive suggesting the presence of an altered component of transcription inside mitochondria. The defect was confirmed by studies with a transcriptionally active DNA-protein complex and by testing the DNA-binding ability of mitochondrial proteins.
在我们先前的工作中(利索夫斯基等人,1987年;利索夫斯基和米夏埃利斯,1988年),我们鉴定出酵母的两个核pet基因,它们是线粒体转录所必需的。在本报告中,我们表明这些pet突变之一,pet-ts798,定位于编码线粒体RNA聚合酶的RP041基因中。当插入高拷贝数质粒时,突变体pet-ts798的温度敏感损伤可被第二个核基因RF1023(mtf1)抑制。我们关于mtf1编码一种40 kDa线粒体转录因子的假设得到了以下事实的支持:克隆基因作为温度敏感RNA聚合酶突变体的基因间抑制子起作用。通过分析突变体pet-ts3504鉴定出第三个线粒体转录核基因(mtf2)。分离的突变线粒体的体外转录活性对温度敏感,这表明线粒体内存在一种改变的转录成分。通过对转录活性DNA-蛋白质复合物的研究以及测试线粒体蛋白质的DNA结合能力,证实了该缺陷。