DNM1, a dynamin-related gene, participates in endosomal trafficking in yeast.
We identified DNM1, a novel dynamin-related gene in Saccharomyces cerevisiae. Molecular and genetic mapping showed that DNM1 is the most proximal gene to the right of centromere 12, and is predicted to encode a protein of 85 kD, designated Dnm1p. The protein exhibits 41% overall identity with full-length dynamin I and 55% identity with the most highly conserved 400-amino acid GTPase region. Our findings show that like mammalian dynamin, Dnm1p participates in endocytosis; however, it is unlikely to be a cognate homologue. Cells with a disruption in the DNM1 gene showed mating response defects consistent with a delay in receptor-mediated endocytosis. The half-life of the Ste3p pheromone receptor was increased two- to threefold in the dnm1 mutant, demonstrating that Dnm1p participates in the constitutive turnover of the receptor. To define the step in the endocytic pathway at which Dnm1p acts, we analyzed mutant strains at both early and late steps of the process. Initial internalization of epitope-tagged pheromone receptor or of labeled pheromone proceeded with wild-type kinetics. However, delivery of the internalized receptor to the vacuole was greatly impeded during ligand-induced endocytosis. These data suggest that during receptor-mediated endocytosis, Dnm1p acts after internalization, but before fusion with the vacuole. The dnm1 mutant was not defective for sorting of vacuolar proteins, indicating that Dnm1p is not required for transport from the late endosome to the vacuole. Therefore, we suggest that Dnm1p participates at a novel step before fusion with the late endosome.
Gammie
A E
AE
Department of Molecular Biology, Princeton University, New Jersey 08544-1014, USA.
Kurihara
L J
LJ
Vallee
R B
RB
Rose
M D
MD
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D003001:Cloning, Molecular; D034284:Dynamin I; D034281:Dynamins; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000235:genetics*; D020558:GTP Phosphohydrolases / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D008024:Ligands; D000072235:Mating Factor; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides / Q000378:metabolism; D010675:Pheromones / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D051964:Receptors, Mating Factor; D043563:Receptors, Pheromone; D012098:Reproduction; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014617:Vacuoles / Q000378:metabolism
D005656:Fungal Proteins; D008024:Ligands; D024101:Mitochondrial Proteins; D010455:Peptides; D010675:Pheromones; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D051964:Receptors, Mating Factor; D043563:Receptors, Pheromone; C081863:STE3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000072235:Mating Factor; D034284:Dynamin I; D020558:GTP Phosphohydrolases; C094774:DNM1 protein, S cerevisiae; D034281:Dynamins