Groudinsky O, Bousquet I, Wallis M G, Slonimski P P, Dujardin G
Centre de Génétique Moléculaire, Laboratoire propre du CNRS associé à l'Université P. et M. Curie, Gif sur Yvette, France.
Mol Gen Genet. 1993 Sep;240(3):419-27. doi: 10.1007/BF00280396.
The NAM1/MTF2 gene was firstly isolated as a multicopy suppressor of mitochondrial splicing deficiencies and independently as a gene of which a thermosensitive allele affects mitochondrial transcription in organello. To determine which step in mitochondrial RNA metabolism is controlled in vivo by the NAM1 gene, mitochondrial transcripts of seven transcription units from strains carrying an inactive nam1::URA3 gene disruption in various mitochondrial genetic backgrounds were analysed by Northern blot hybridisations. In a strain carrying an intron-containing mitochondrial genome, the inactivation of the NAM1 gene led to a strong decrease in (or total absence of) the mosaic cytb and cox1 mRNAs and in transcripts of the atp6-rf3/ens2 genes, which are co-transcribed with cox1. Neither the accumulation of unspliced cytb or cox1 pre-mRNAs, nor that of excised circular intron molecules of ai1 or ai2 were observed, but the abundance of the bi1 and ai7 lariats was comparable to that observed in the wild-type strain, thus demonstrating that transcription of the cytb and cox1 genes does occur. In strains carrying the intron-less mitochondrial genome with or without the rf3/ens2 sequence, wild-type amounts of cytb and cox1 mRNAs were detected while the amount of the atp6 mRNA was always strongly decreased. The abundance of transcripts from five other genes was either slightly (21S rRNA) or not at all (cox2, cox3, atp9 and 15S rRNA) affected by the nam1 inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)
NAM1/MTF2基因最初是作为线粒体剪接缺陷的多拷贝抑制因子被分离出来的,并且独立地作为一个基因,其温度敏感等位基因会影响细胞器中的线粒体转录。为了确定线粒体RNA代谢的哪个步骤在体内受NAM1基因控制,通过Northern印迹杂交分析了在各种线粒体遗传背景下携带无活性nam1::URA3基因破坏的菌株中七个转录单元的线粒体转录本。在携带含内含子线粒体基因组的菌株中,NAM1基因的失活导致镶嵌型细胞色素b(cytb)和细胞色素c氧化酶亚基1(cox1)mRNA以及与cox1共同转录的atp6-rf3/ens2基因转录本大幅减少(或完全缺失)。未观察到未剪接的cytb或cox1前体mRNA的积累,也未观察到ai1或ai2切除的环状内含子分子的积累,但bi1和ai7套索状结构的丰度与野生型菌株中观察到的相当,从而证明cytb和cox1基因的转录确实发生。在携带有无rf3/ens2序列的无内含子线粒体基因组的菌株中,检测到野生型水平的cytb和cox1 mRNA,而atp6 mRNA的量总是大幅减少。nam1失活对其他五个基因转录本的丰度影响轻微(21S rRNA)或没有影响(cox2、cox3、atp9和15S rRNA)。(摘要截短于250字)