Lu Lin, Roberts George, Simon Kirk, Yu Jia, Hudson Alan P
Department of Immunology and Microbiology, Wayne State University School of Medicine, Gordon H. Scott Hall, 540 East Canfield Avenue, Detroit, MI 48201, USA.
Curr Genet. 2003 Jul;43(4):263-72. doi: 10.1007/s00294-003-0398-z. Epub 2003 May 7.
A central problem in our understanding of mitochondrial (mt) function remains the question of how coordinate transcriptional control is accomplished between nucleus and mitochondria. Here, we report the initial characterization of a protein of previously unknown function, the product of the YMR030 W gene, that appears to mediate such coordinate gene expression. Expression of YMR030 W is glucose-repressible; a deletion mutant for this gene shows a severe growth defect on glycerol-, but not glucose- or ethanol-based medium. In that mutant, transcript levels from GUT1 and GUT2 are highly attenuated compared with those of the wild-type parent when both are grown on glycerol-based medium. Under the same growth conditions, transcripts from the mt OLI1 gene, which has one copy of a mt upstream activating sequence (UAS) in its 5'-flanking region, are attenuated in the DeltaYMR030 W mutant, but mRNA from the mt COX3 ( OXI2) gene, which lacks the mt UAS, are not. Some nuclear genes encoding mt-related proteins also show low transcript levels in the DeltaYMR030 W mutant in comparison with those of the wild-type parent strain during glycerol-based growth. Localization of the protein, via its expression fused to green fluorescent protein, indicates that it is present in both nucleus and mitochondria, supporting a respiration-related transcriptional role for this gene product in both cellular genetic compartments. Because of its role in both respiratory growth and mt function, we designate the YMR030 W coding sequence RSF1 (respiration factor 1).
我们在理解线粒体(mt)功能方面的一个核心问题仍然是,如何在细胞核和线粒体之间实现协调转录控制。在此,我们报道了一种功能此前未知的蛋白质(YMR030W基因的产物)的初步特征,该蛋白质似乎介导了这种协调基因表达。YMR030W的表达受葡萄糖抑制;该基因的缺失突变体在基于甘油的培养基上表现出严重的生长缺陷,但在基于葡萄糖或乙醇的培养基上则没有。在该突变体中,当在基于甘油的培养基上生长时,与野生型亲本相比,GUT1和GUT2的转录水平大幅降低。在相同的生长条件下,mt OLI1基因(其5'侧翼区域有一个mt上游激活序列(UAS)拷贝)的转录本在DeltaYMR030W突变体中减少,但缺乏mt UAS的mt COX3(OXI2)基因的mRNA则没有减少。一些编码与mt相关蛋白质的核基因在基于甘油生长期间,与野生型亲本菌株相比,在DeltaYMR030W突变体中也显示出低转录水平。通过与绿色荧光蛋白融合表达对该蛋白质进行定位,表明它同时存在于细胞核和线粒体中,这支持了该基因产物在两个细胞遗传区室中具有与呼吸相关的转录作用。由于其在呼吸生长和mt功能中的作用,我们将YMR030W编码序列命名为RSF1(呼吸因子1)。