Abstract
Cdc50p, a transmembrane protein localized to the late endosome, is required for polarized cell growth in yeast. Genetic studies suggest that CDC50 performs a function similar to DRS2, which encodes a P-type ATPase of the aminophospholipid translocase (APT) subfamily. At low temperatures, drs2Delta mutant cells exhibited depolarization of cortical actin patches and mislocalization of polarity regulators, such as Bni1p and Gic1p, in a manner similar to the cdc50Delta mutant. Both Cdc50p and Drs2p were localized to the trans-Golgi network and late endosome. Cdc50p was coimmunoprecipitated with Drs2p from membrane protein extracts. In cdc50Delta mutant cells, Drs2p resided on the endoplasmic reticulum (ER), whereas Cdc50p was found on the ER membrane in drs2Delta cells, suggesting that the association on the ER membrane is required for transport of the Cdc50p-Drs2p complex to the trans-Golgi network. Lem3/Ros3p, a homolog of Cdc50p, was coimmunoprecipitated with another APT, Dnf1p; Lem3p was required for exit of Dnf1p out of the ER. Both Cdc50p-Drs2p and Lem3p-Dnf1p were confined to the plasma membrane upon blockade of endocytosis, suggesting that these proteins cycle between the exocytic and endocytic pathways, likely performing redundant functions. Thus, phospholipid asymmetry plays an important role in the establishment of cell polarity; the Cdc50p/Lem3p family likely constitute potential subunits specific to unique P-type ATPases of the APT subfamily.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP-Binding Cassette Transporters
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Adaptor Proteins, Signal Transducing
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Adenosine Triphosphatases / analysis
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Adenosine Triphosphatases / metabolism
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Calcium-Transporting ATPases / analysis
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Calcium-Transporting ATPases / genetics
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Calcium-Transporting ATPases / metabolism*
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Carrier Proteins / analysis
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Carrier Proteins / metabolism
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Cell Membrane / chemistry
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Cell Membrane / metabolism
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Cell Polarity
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Endosomes / chemistry
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Endosomes / metabolism
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Membrane Proteins / genetics
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Membrane Transport Proteins / analysis
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Membrane Transport Proteins / metabolism
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Microfilament Proteins / analysis
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Microfilament Proteins / metabolism
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Phospholipid Transfer Proteins
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Phospholipids / metabolism*
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins / analysis
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Sequence Deletion / genetics
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trans-Golgi Network / chemistry
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trans-Golgi Network / metabolism
Substances
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ATP-Binding Cassette Transporters
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Adaptor Proteins, Signal Transducing
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Bni1 protein, S cerevisiae
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CDC50 protein, S cerevisiae
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Carrier Proteins
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DRS2 protein, S cerevisiae
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GIC1 protein, S cerevisiae
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Lem3 protein, S cerevisiae
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Membrane Proteins
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Membrane Transport Proteins
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Microfilament Proteins
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Phospholipid Transfer Proteins
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Phospholipids
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Saccharomyces cerevisiae Proteins
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Adenosine Triphosphatases
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Calcium-Transporting ATPases
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Dnf1 protein, S cerevisiae
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NEO1 protein, S cerevisiae