The sequence NPFXD defines a new class of endocytosis signal in Saccharomyces cerevisiae
| Autor: | Gregory S. Payne, J P Howard, Philip K. Tan |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Receptors Peptide Lipoproteins Recombinant Fusion Proteins Endocytic cycle Golgi Apparatus Saccharomyces cerevisiae Protein Sorting Signals Biology Endocytosis Pheromones Exocytosis Fungal Proteins symbols.namesake Bacterial Proteins Amino Acid Sequence Subtilisins Golgi localization Golgi membrane Lysine Helix-Loop-Helix Motifs Biological Transport Articles Cell Biology Receptor-mediated endocytosis Golgi apparatus Immunohistochemistry Clathrin Cell biology DNA-Binding Proteins Repressor Proteins Cytoskeletal Proteins Receptors Mating Factor LDL receptor symbols Tyrosine Proprotein Convertases Transcription Factors |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| DOI: | 10.1083/jcb.135.6.1789 |
| Popis: | The yeast membrane protein Kex2p uses a tyrosine-containing motif within the cytoplasmic domain for localization to a late Golgi compartment. Because Golgi membrane proteins mislocalized to the plasma membrane in yeast can undergo endocytosis, we examined whether the Golgi localization sequence or other sequences in the Kex2p cytoplasmic domain mediate endocytosis. To assess endocytic function, the Kex2p cytoplasmic domain was fused to an endocytosis-defective form of the alpha-factor receptor. Ste2p. Like intact Ste2p, the chimeric protein, Stex22p, undergoes rapid endocytosis that is dependent on clathrin and End3p. Uptake of Stex22p does not require the Kex2p Golgi localization motif. Instead, the sequence NPFSD, located 37 amino acids from the COOH terminus, is essential for Stex22p endocytosis. Internalization was abolished when the N, P, or F residues were converted to alanine and severely impaired upon conversion of D to A. NPFSD restored uptake when added to the COOH terminus of an endocytosis-defective Ste2p chimera lacking lysine-based endocytosis signals present in wild-type Ste2p. An NPF sequence is present in the cytoplasmic domain of the a-factor receptor, Ste3p. Mutation of this sequence prevented pheromone-stimulated endocytosis of a truncated form of Ste3p. Our results identify NPFSD as a clathrin-dependent endocytosis signal that is distinct from the aromatic amino acid-containing Golgi localization motif and lysine-based, ubiquitin-dependent endocytosis signals in yeast. |
| Databáze: | OpenAIRE |
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