Acidification of the lysosome-like vacuole and the vacuolar H+-ATPase are deficient in two yeast mutants that fail to sort vacuolar proteins
Acidification of the lysosome-like vacuole and the vacuolar H+-ATPase are deficient in two yeast mutants that fail to sort vacuolar proteins
复制标题
两种无法分选液泡蛋白的酵母突变体中，溶酶体样液泡的酸化和液泡 H-ATP 酶存在缺陷
作者：
J. H. Rothman;Carl T. Yamashiro;C K Raymond;Patricia M. Kane;T. Stevens
细胞器酸化在动物细胞的细胞内蛋白质加工、运输和分选中起着明显的作用。我们研究了酸化和蛋白质分选在酵母中的关系，通过处理酵母细胞与氯化铵，发现这种lysosomotropic剂引起的错误定位的一个新合成的空泡（溶酶体）酶蛋白酶A（PrA）的细胞表面的相当大的一部分。我们还确定了vpl突变体的一个亚组，其在液泡蛋白的分选中有缺陷（Rothman，J.H.，和T. H.史蒂文斯1986. Cell. 47：1041-1051; Rothman，J.H.，I. Howald和T. H.史蒂文斯EMBO [Eur.摩尔生物器官] J.在出版中），未能在其空泡内积累亲溶酶体荧光染料奎纳克林，模拟用铵处理的野生型细胞的表型。vp 13和vp 16突变体的酸化缺陷与液泡ATP酶活性的显著缺陷、纯化的液泡膜中质子转运ATP酶（H+-ATP酶）的两个免疫反应性亚基的水平降低以及PrA作为前体物质的细胞内部分的积累相关。因此，一些VPL基因是酵母液泡H+-ATP酶复合物正常功能所必需的，并且可以编码酶的亚基或其组装和靶向所需的组分。总的来说，这些研究结果暗示了酸化在空泡蛋白分选和酵母酶原激活中的关键作用，并表明酵母空泡酸化系统的组成部分可以通过检查空泡蛋白分选缺陷的突变体来确定。
Organelle acidification plays a demonstrable role in intracellular protein processing, transport, and sorting in animal cells. We investigated the relationship between acidification and protein sorting in yeast by treating yeast cells with ammonium chloride and found that this lysosomotropic agent caused the mislocalization of a substantial fraction of the newly synthesized vacuolar (lysosomal) enzyme proteinase A (PrA) to the cell surface. We have also determined that a subset of the vpl mutants, which are deficient in sorting of vacuolar proteins (Rothman, J. H., and T. H. Stevens. 1986. Cell. 47:1041-1051; Rothman, J. H., I. Howald, and T. H. Stevens. EMBO [Eur. Mol. Biol. Organ.] J. In press), failed to accumulate the lysosomotropic fluorescent dye quinacrine within their vacuoles, mimicking the phenotype of wild-type cells treated with ammonium. The acidification defect of vpl3 and vpl6 mutants correlated with a marked deficiency in vacuolar ATPase activity, diminished levels of two immunoreactive subunits of the protontranslocating ATPase (H+-ATPase) in purified vacuolar membranes, and accumulation of the intracellular portion of PrA as the precursor species. Therefore, some of the VPL genes are required for the normal function of the yeast vacuolar H+-ATPase complex and may encode either subunits of the enzyme or components required for its assembly and targeting. Collectively, these findings implicate a critical role for acidification in vacuolar protein sorting and zymogen activation in yeast, and suggest that components of the yeast vacuolar acidification system may be identified by examining mutants defective in sorting of vacuolar proteins.
中国仓鼠细胞突变体中的热敏损伤对内体和溶酶体的酸化产生不同的影响。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Timchak,LM;Kruse,F;Marnell,MH;Draper,RK
通讯作者：
Draper,RK
来自酸化缺陷的中国仓鼠细胞系的网格蛋白包被的囊泡组分中不耐热质子易位 ATP 酶和泵活性。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Stone,DK;Marnell,M;Yang,Y;Draper,RK
通讯作者：
Draper,RK
DOI：
10.1073/pnas.85.21.7972
发表时间：
1988-11-01
影响因子：
11.1
作者：
BOWMAN, EJ;SIEBERS, A;ALTENDORF, K
通讯作者：
ALTENDORF, K