A novel immunodetection screen for vacuolar defects identifies a unique allele of VPS35 in S. cerevisiae.
A novel immunodetection screen for vacuolar defects identifies a unique allele of VPS35 in S. cerevisiae.
复制标题
一种针对液泡缺陷的新型免疫检测筛选鉴定了酿酒酵母中 VPS35 的独特等位基因。
DOI：
10.1007/s11010-008-9703-y
复制
发表时间：
2008
影响因子：
4.3
通讯作者：
Gharakhanian,Editte
中科院分区：
文献类型：
--
作者：
Takahashi,MKathleen;Frost,Christopher;Oyadomari,Ken;Pinho,Marcos;Sao,Dyna;Chima-Okereke,Onyi;Gharakhanian,Editte
The late endosome and vacuole of yeastSaccharomyces cerevisiaeare functionally equivalent to the mammalian late endosome and lysosome. The late endosome is the convergence point of the biosynthetic and endocytic trafficking to the vacuole. Here, we describe a novel immunodetection screen to isolate mutants defective in trafficking the soluble hydrolase carboxypeptidase Y (CPY) at the late endosome to vacuole interface (envmutants). Mutants exhibit vacuolar morphology and endocytosis defects as assayed by electron, fluorescent, and nomarski microscopy. In biochemical assays, they internally accumulate p2CPY in a dense membrane compartment lacking vacuolar properties yet display normal secretion phenotypes. The results suggest vacuolar morphology and function defects that are exclusively at the late endosome/vacuole interface.envmutants define five complementation groups. The first gene of the collection to be cloned,ENV1is allelic toVPS35whose established function is in retrograde trafficking from late endosome to trans-Golgi network (TGN). Microscopic, biochemical, and growth analyses establish thatenv1is distinct from other alleles ofVPS35in vacuolar morphology, growth characteristics, and internal accumulation of p2CPY. Our results indicate thatENVgenes may define new gene functions at the late endosome to vacuole interface.
