Molecular analysis of the yeast VPS3 gene and the role of its product in vacuolar protein sorting and vacuolar segregation during the cell cycle.
Molecular analysis of the yeast VPS3 gene and the role of its product in vacuolar protein sorting and vacuolar segregation during the cell cycle.
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DOI：
10.1083/jcb.111.3.877
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发表时间：
1990-09
期刊：
影响因子：
--
通讯作者：
Stevens TH
中科院分区：
文献类型：
--
作者：
Raymond CK;O'Hara PJ;Eichinger G;Rothman JH;Stevens TH
酿酒酵母的vps 3突变体在新合成的可溶性液泡蛋白的分选和液泡的酸化中受损（Rothman，J.H.，和T. H.史蒂文斯Cell. 47：1041-1051; Rothman，J.H.，C. T.山城角K.雷蒙德，P.M. Kane和T. H.史蒂文斯1989. 109：93-100）。VPS 3基因，这是克隆使用一种新的选择程序，编码低丰度，亲水性蛋白质的117 kD，最有可能驻留在细胞质中。携带VPS 3基因缺失（vps 3- delta 1）的酵母菌株是活的，但它们的生长速率相对于野生型细胞显著降低。用携带温度条件vps 3等位基因的菌株进行的温度变化实验表明，细胞在VPS 3功能丧失后迅速丧失对液泡蛋白羧肽酶Y进行分选的能力。通过荧光显微镜检查野生型和vps 3-delta 1酵母菌株中的酵母细胞形态。野生型酵母细胞中的空泡在形态上是复杂的，并且在芽生长的早期阶段，它们似乎在母细胞和芽之间被积极地分配。vps 3-delta 1突变体中的液泡形态与野生型模式相比发生了显著改变，并且在野生型菌株中观察到的液泡分离过程在这些突变体中是有缺陷的。除了空泡酸化缺陷外，vps 3-delta 1菌株的表型与缺乏空泡质子转运ATP酶的突变体的表型显著不同。这些数据表明vps 3-delta 1细胞中的酸化缺陷不是在这些突变体中观察到的液泡功能多效性缺陷的主要原因。
vps3 mutants of the yeast Saccharomyces cerevisiae are impaired in the sorting of newly synthesized soluble vacuolar proteins and in the acidification of the vacuole (Rothman, J. H., and T. H. Stevens. Cell. 47:1041-1051; Rothman, J. H., C. T. Yamashiro, C. K. Raymond, P. M. Kane, and T. H. Stevens. 1989. J. Cell Biol. 109:93-100). The VPS3 gene, which was cloned using a novel selection procedure, encodes a low abundance, hydrophilic protein of 117 kD that most likely resides in the cytoplasm. Yeast strains bearing a deletion of the VPS3 gene (vps3- delta 1) are viable, yet their growth rate is significantly reduced relative to wild-type cells. Temperature shift experiments with strains carrying a temperature conditional vps3 allele demonstrate that cells rapidly lose the capacity to sort the vacuolar protein carboxypeptidase Y upon loss of VPS3 function. Vacuolar morphology was examined in wild- type and vps3-delta 1 yeast strains by fluorescence microscopy. The vacuoles in wild-type yeast cells are morphologically complex, and they appear to be actively partitioned between mother cells and buds during an early phase of bud growth. Vacuolar morphology in vps3-delta 1 mutants is significantly altered from the wild-type pattern, and the vacuolar segregation process seen in wild-type strains is defective in these mutants. With the exception of a vacuolar acidification defect, the phenotypes of vps3-delta 1 strains are significantly different from those of mutants lacking the vacuolar proton-translocating ATPase. These data demonstrate that the acidification defect in vps3-delta 1 cells is not the primary cause of the pleiotropic defects in vacuolar function observed in these mutants.