改变Mod5p线粒体-细胞质分布的突变表明肌动蛋白细胞骨架以及mRNA 3'末端和/或蛋白质合成参与线粒体运输。
Mutations altering the mitochondrial-cytoplasmic distribution of Mod5p implicate the actin cytoskeleton and mRNA 3' ends and/or protein synthesis in mitochondrial delivery.
作者信息
Zoladek T, Vaduva G, Hunter L A, Boguta M, Go B D, Martin N C, Hopper A K
机构信息
Department of Biochemistry and Molecular Biology, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033, USA.
出版信息
Mol Cell Biol. 1995 Dec;15(12):6884-94. doi: 10.1128/MCB.15.12.6884.
The Saccharomyces cerevisiae MOD5 gene encodes proteins that function in three subcellular locations: mitochondria, the cytoplasm, and nuclei (M. Boguta, L.A. Hunter, W.-C. Shen, E. C. Gillman, N. C. Martin, and A. K. Hopper, Mol. Cell. Biol. 14:2298-2306, 1994; E. C. Gillman, L. B. Slusher, N. C. Martin, and A. K. Hopper, Mol. Cell. Biol. 11:2382-2390, 1991). A mutant allele of MOD5 encoding a protein (Mod5p-I,KR6) located predominantly in mitochondria was constructed. Mutants defective in delivering Mod5p-I,KR6 to mitochondria were sought by selecting cells with increased cytosolic activity of this protein. Twenty-five mutants defining four complementation groups, mdp1, mdp2, mdp3, and mdp4, were found. They are unable to respire at 34 degrees C or to grow on glucose medium at 38 degrees C. Cell fractionation studies showed that mdp1, mdp2, and mdp3 mutants have an altered mitochondrial-cytoplasmic distribution of Mod5p. mdp2 can be suppressed by ACT1, the actin-encoding gene. The actin cytoskeleton organization is also aberrant in mdp2 cells. MDP2 is the same as VRP1 (S. F. H. Donnelly, M. J. Picklington, D. Pallotta, and E. Orr, Mol. Microbiol. 10:585-596, 1993). MDP3 is identical to PAN1, which encodes a protein that interacts with mRNA 3' ends and affects initiation of protein synthesis (A. B. Sachs and J. A. Deardoff, Cell 70:961-973, 1992). These results implicate the actin cytoskeleton and mRNA 3' ends and/or protein synthesis as being as important for protein distribution in S. cerevisiae as they are for distribution of cytosolic proteins in higher eukaryotes. This provides the potential to apply genetic and molecular approaches to study gene products and mechanisms involved in this type of protein distribution. The selection strategy also offers a new approach for identifying gene products involved in the distribution of proteins to their subscellular destinations.
酿酒酵母的MOD5基因编码的蛋白质在三个亚细胞位置发挥作用：线粒体、细胞质和细胞核（M. 博古塔、L.A. 亨特、W.-C. 沈、E.C. 吉尔曼、N.C. 马丁和A.K. 霍珀，《分子细胞生物学》14:2298 - 2306，1994；E.C. 吉尔曼、L.B. 斯拉舍、N.C. 马丁和A.K. 霍珀，《分子细胞生物学》11:2382 - 2390，1991）。构建了一个MOD5的突变等位基因，其编码一种主要定位于线粒体的蛋白质（Mod5p - I,KR6）。通过筛选该蛋白质胞质活性增加的细胞，寻找将Mod5p - I,KR6转运至线粒体存在缺陷的突变体。发现了25个定义了四个互补群mdp1、mdp2、mdp3和mdp4的突变体。它们在34℃下无法进行呼吸作用，在38℃下无法在葡萄糖培养基上生长。细胞分级分离研究表明，mdp1、mdp2和mdp3突变体中Mod5p的线粒体 - 细胞质分布发生了改变。mdp2可以被ACT1（肌动蛋白编码基因）抑制。mdp2细胞中的肌动蛋白细胞骨架组织也异常。MDP2与VRP1相同（S.F.H. 唐纳利、M.J. 皮克林顿、D. 帕洛塔和E. 奥尔，《分子微生物学》10:585 - 596，1993）。MDP3与PAN1相同，PAN1编码一种与mRNA 3' 末端相互作用并影响蛋白质合成起始的蛋白质（A.B. 萨克斯和J.A. 迪尔多夫，《细胞》70:961 - 973，1992）。这些结果表明，肌动蛋白细胞骨架以及mRNA 3' 末端和/或蛋白质合成对于酿酒酵母中的蛋白质分布与它们对于高等真核生物中胞质蛋白质的分布一样重要。这为应用遗传和分子方法研究参与此类蛋白质分布的基因产物和机制提供了可能性。这种筛选策略还为鉴定参与蛋白质向其亚细胞目的地分布的基因产物提供了一种新方法。
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