De Haan M, van Loon A P, Kreike J, Vaessen R T, Grivell L A
Eur J Biochem. 1984 Jan 2;138(1):169-77. doi: 10.1111/j.1432-1033.1984.tb07896.x.
The nuclear gene coding for the imported 14-kDa subunit of the ubiquinol-cytochrome c reductase of yeast mitochondria has been sequenced in an attempt to define regulatory and protein topogenic elements. The gene has a length of 381 base pairs and is potentially capable of encoding a polypeptide of 14561 Da. It is transcribed into a single low-abundance RNA of 680 nucleotides whose 5' and 3' termini map, respectively, 30-35 nucleotides upstream and 180-190 nucleotides downstream of the initiator and termination codons. Consistent with the estimated low level of the mRNA, codon usage in the gene is not strongly biased and other features, characteristic of highly expressed genes in yeast, are absent. The 14-kDa protein is predicted to be a predominantly hydrophilic protein, with only a single, short hydrophobic stretch located between positions 19-38. Comparison with other imported mitochondrial proteins so far sequenced has failed to reveal unifying features that might serve as targeting elements. Steady-state levels of the 14-kDa and 11-kDa subunits are reduced in mit- mutants which synthesize truncated forms of apocytochrome b and in these, newly synthesized subunits exhibit a specifically increased turnover rate. We suggest that association of these two subunits with the complex may be mediated or enhanced by interaction with other subunits, in particular cytochrome b.
为了确定调控元件和蛋白质拓扑结构元件,对酵母线粒体泛醇 - 细胞色素c还原酶中导入的14 kDa亚基的核基因进行了测序。该基因长度为381个碱基对,理论上能够编码一个14561 Da的多肽。它转录成一种680个核苷酸的低丰度单链RNA,其5'和3'末端分别位于起始密码子和终止密码子上游30 - 35个核苷酸以及下游180 - 190个核苷酸处。与估计的低水平mRNA一致,该基因的密码子使用没有强烈偏向性,并且缺乏酵母中高表达基因所特有的其他特征。预测14 kDa蛋白主要是亲水性蛋白,仅在19 - 38位之间有一个单一的短疏水区域。与目前已测序的其他导入线粒体蛋白进行比较,未能揭示可能作为靶向元件的统一特征。在合成截短形式脱辅基细胞色素b的mit - 突变体中,14 kDa和11 kDa亚基的稳态水平降低,并且在这些突变体中,新合成的亚基表现出特异性增加的周转率。我们认为这两个亚基与复合物的结合可能是由与其他亚基(特别是细胞色素b)的相互作用介导或增强的。