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An Overexpression Screen in Saccharomyces cerevisiae Identifies Novel Genes that Affect Endocytic Protein Trafficking
Author(s) -
Arlt Henning,
Perz Angela,
Ungermann Christian
Publication year - 2011
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/j.1600-0854.2011.01252.x
Subject(s) - biology , endocytic cycle , saccharomyces cerevisiae , microbiology and biotechnology , gene , affect (linguistics) , yeast , transport protein , genetics , endocytosis , cell , linguistics , philosophy
A large number of proteins involved in the biogenesis of yeast endosomes and vacuoles have been identified based on screens that scored for inactivation of proteins. Such screens may, however, miss important regulators of the pathway. Here, we present a visual screen in which we examined the effects on vacuole morphology if any of the 6153 yeast open reading frames was overexpressed. Using a progressive screening procedure, we could identify a total of 53 genes. Among the most striking endosomal proteins are the CORVET/HOPS subunits Vps3, Vps18 and Vps39 and the putative tethering inhibitor Ivy1. Furthermore, six endosomal sorting complex related to transport (ESCRT) proteins led to altered vacuole morphology if overproduced. Among the novel proteins, we identify Yer128w as an endosomal protein that interacts with the AAA‐ATPase Vps4, and therefore named it Vfa1 ( V ps F our‐ A ssociated 1). We present evidence on the possible role of these novel proteins in trafficking to the vacuole. Our data provide novel insights into the regulation of protein trafficking.