The Ypt1 GTPase is essential for the first two steps of the yeast secretory pathway.
Small GTPases of the rab family are involved in the regulation of vesicular transport. The restricted distribution of each of these proteins in mammalian cells has led to the suggestion that different rab proteins act at different steps of transport (Pryer, N. K., L. J. Wuestehube, and R. Sheckman. 1992. Annu Rev. Biochem. 61:471-516; Zerial, M., and H. Stenmark. 1993. Curr. Opin. Cell Biol. 5:613-620). However, in this report we show that the Ypt1-GTPase, a member of the rab family, is essential for more than one step of the yeast secretory pathway. We determined the secretory defect conferred by a novel ypt1 mutation by comparing the processing of several transported glycoproteins in wild-type and mutant cells. The ypt1-A136D mutant has a change in an amino acid that is conserved among rab GTPases. This mutation leads to a rapid and tight secretory block upon a shift to the restrictive temperature, and allows for the identification of the specific steps in the secretory pathway that directly require Ypt1 protein (Ypt1p). The ypt1-A136D mutant exhibits tight blocks in two secretory steps, ER to cis-Golgi and cis- to medial-Golgi, but later steps are unaffected. Thus, it is unlikely that Ypt1p functions as the sole determinant of fusion specificity. Our results are more consistent with a role for Ypt1/rab proteins in determining the directionality or fidelity of protein sorting.
Jedd
G
G
Department of Pharmacological and Physiological Sciences, University of Chicago, Illinois 60637, USA.
Richardson
C
C
Litt
R
R
Segev
N
N
D001483:Base Sequence; D003594:Cytoplasmic Granules / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology; D010641:Phenotype; D029701:Saccharomyces cerevisiae Proteins; D013696:Temperature; D015003:Yeasts / Q000502:physiology*; D020691:rab GTP-Binding Proteins
D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; C060755:YPT1 protein, S cerevisiae; D020691:rab GTP-Binding Proteins