Defects in structural integrity of ergosterol and the Cdc50p-Drs2p putative phospholipid translocase cause accumulation of endocytic membranes, onto which actin patches are assembled in yeast.
Specific changes in membrane lipid composition are implicated in actin cytoskeletal organization, vesicle formation, and control of cell polarity. Cdc50p, a membrane protein in the endosomal/trans-Golgi network compartments, is a noncatalytic subunit of Drs2p, which is implicated in translocation of phospholipids across lipid bilayers. We found that the cdc50Delta mutation is synthetically lethal with mutations affecting the late steps of ergosterol synthesis (erg2 to erg6). Defects in cell polarity and actin organization were observed in the cdc50Delta erg3Delta mutant. In particular, actin patches, which are normally found at cortical sites, were assembled intracellularly along with their assembly factors, including Las17p, Abp1p, and Sla2p. The exocytic SNARE Snc1p, which is recycled by an endocytic route, was also intracellularly accumulated, and inhibition of endocytic internalization suppressed the cytoplasmic accumulation of both Las17p and Snc1p. Simultaneous loss of both phospholipid asymmetry and sterol structural integrity could lead to accumulation of endocytic intermediates capable of initiating assembly of actin patches in the cytoplasm.
Kishimoto
Takuma
T
Division of Molecular Interaction, Institute for Genetic Medicine, Hokkaido University Graduate School of Medicine, N15 W7, Kita-ku, Sapporo 060-0815, Japan.
Yamamoto
Takaharu
T
Tanaka
Kazuma
K
D000199:Actins / Q000096:biosynthesis*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D016764:Cell Polarity; D004705:Endocytosis / Q000502:physiology*; D004875:Ergosterol / Q000494:pharmacology*; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D005838:Genotype; D007425:Intracellular Membranes / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D000199:Actins; C427246:CDC50 protein, S cerevisiae; C104014:DRS2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000252:Calcium-Transporting ATPases; D004875:Ergosterol