Wiesenberger Gerlinde, Speer Falk, Haller Günter, Bonnefoy Nathalie, Schleiffer Alexander, Schafer Bernd
Max F. Perutz Laboratories, Department of Genetics, University of Vienna, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria.
J Mol Biol. 2007 Mar 30;367(3):681-91. doi: 10.1016/j.jmb.2007.01.014. Epub 2007 Jan 10.
We report here on the role of open reading frame (ORF) SPCC1183.04c of Schizosaccharomyces pombe in mitochondrial RNA metabolism. A mutant deleted for this ORF on chromosome III accumulates mitochondrial transcripts with the exception of the cob mRNA. A detailed Northern blot analysis showed that the effect results from a decrease in RNA degradation but not from RNA processing deficiencies. Overexpression of the SPCC1183.04c gene in a S. pombe wild-type strain is characterized by slow growth at 37 degrees C on non-fermentable carbon sources and a significant reduction of steady-state levels of mitochondrial transcripts. A NCBI BLASTP search with the amino acid sequence deduced from the S. pombe gene identified significant similarity to a number of proteins in fungi (e.g. Ascomycota, Basidiomycota) and in some non-fungal eukaryotes (e.g. ciliate, slime mold, red algae). By heterologous expression of SPCC1183.04c in a Saccharomyces cerevisiae pet127Delta strain, we demonstrate that the fission yeast protein and Pet127p from S. cerevisiae function similarly: The fission yeast gene complemented the respiratory defect associated with the pet127Delta allele and partially restored the RNA processing phenotype. Although it lacks any recognizable targeting signal, the S. pombe protein is imported into S. cerevisiae mitochondria in vivo. We conclude from our results that the fission yeast SPCC1183.04c gene is a member of a new protein family that functions to stimulate mitochondrial RNA degradation, a function that is conserved within the mitochondria of lower eukaryotes but seems to have been replaced by alternative pathways in metazoans and higher plants.
我们在此报告粟酒裂殖酵母开放阅读框(ORF)SPCC1183.04c在线粒体RNA代谢中的作用。在第三条染色体上缺失该ORF的突变体除了cob mRNA外,会积累线粒体转录本。详细的Northern印迹分析表明,这种效应是由RNA降解减少而非RNA加工缺陷导致的。在粟酒裂殖酵母野生型菌株中过表达SPCC1183.04c基因的特征是,在37摄氏度的非发酵碳源上生长缓慢,并且线粒体转录本的稳态水平显著降低。使用从粟酒裂殖酵母基因推导的氨基酸序列进行NCBI BLASTP搜索,发现与真菌(如子囊菌门、担子菌门)和一些非真菌真核生物(如纤毛虫、黏菌、红藻)中的许多蛋白质具有显著相似性。通过在酿酒酵母pet127Delta菌株中异源表达SPCC1183.04c,我们证明裂殖酵母蛋白与酿酒酵母的Pet127p功能相似:裂殖酵母基因弥补了与pet127Delta等位基因相关的呼吸缺陷,并部分恢复了RNA加工表型。尽管粟酒裂殖酵母蛋白缺乏任何可识别的靶向信号,但在体内它能被导入酿酒酵母线粒体。我们从结果中得出结论,裂殖酵母SPCC1183.04c基因是一个新的蛋白质家族的成员,该家族的功能是刺激线粒体RNA降解,这种功能在低等真核生物的线粒体中是保守的,但在后生动物和高等植物中似乎已被替代途径所取代。