The ABC-transporter Ste6 Accumulates in the Plasma Membrane in a Ubiquitinated Form in Endocytosis Mutants

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1994 Jul 15

PMID 8045256

Citations 113

Authors

R Kolling

C P Hollenberg

Affiliations

Soon will be listed here.

Abstract

We are investigating the transport and turnover of the multispanning membrane protein Ste6. The Ste6 protein is a member of the ABC-transporter family and is required for the secretion of the yeast mating pheromone a-factor. In contrast to the prevailing view that Ste6 is a plasma membrane protein, we found that Ste6 is mainly associated with internal membranes and not with the cell surface. Fractionation and immunofluorescence data are compatible with a Golgi localization of Ste6. Despite its mostly intracellular localization, the Ste6 protein is in contact with the cell surface, as demonstrated by the finding that Ste6 accumulates in the plasma membrane in endocytosis mutants. The Ste6 protein which accumulates in the plasma membrane in endocytosis mutants is ubiquitinated. Ste6 is thus the second protein in yeast besides MAT alpha 2 for which ubiquitination has been demonstrated. Ste6 is a very unstable protein (half-life 13 min) which is stabilized approximately 3-fold in a ubc4 ubc5 mutant, implicating the ubiquitin system in the degradation of Ste6. The strongest stabilizing effect on Ste6 is, however, observed in the vacuolar pep4 mutant (half-life > 2 h), suggesting that most of Ste6 is degraded in the vacuole. Secretory functions are required for efficient degradation of Ste6, indicating that Ste6 enters the secretory pathway and is transported to the vacuole by vesicular carriers.

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References

Kolling R, Nguyen T, Chen E, Botstein D . A new yeast gene with a myosin-like heptad repeat structure. Mol Gen Genet. 1993; 237(3):359-69. DOI: 10.1007/BF00279439. View

Johnston M, Davis R . Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984; 4(8):1440-8. PMC: 368932. DOI: 10.1128/mcb.4.8.1440-1448.1984. View

Kuchler K, Dohlman H, Thorner J . The a-factor transporter (STE6 gene product) and cell polarity in the yeast Saccharomyces cerevisiae. J Cell Biol. 1993; 120(5):1203-15. PMC: 2119724. DOI: 10.1083/jcb.120.5.1203. View

Rose M, Novick P, Thomas J, Botstein D, Fink G . A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987; 60(2-3):237-43. DOI: 10.1016/0378-1119(87)90232-0. View

Franzusoff A, Rothblatt J, Schekman R . Analysis of polypeptide transit through yeast secretory pathway. Methods Enzymol. 1991; 194:662-74. DOI: 10.1016/0076-6879(91)94048-h. View

Yarden Y, Escobedo J, Kuang W, Yang-Feng T, Daniel T, Tremble P . Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986; 323(6085):226-32. DOI: 10.1038/323226a0. View

Denning G, Anderson M, Amara J, Marshall J, Smith A, Welsh M . Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive. Nature. 1992; 358(6389):761-4. DOI: 10.1038/358761a0. View

Kellermann E, Hollenberg C . The glucose-and ethanol-dependent regulation of PDC1 from Saccharomyces cerevisiae are controlled by two distinct promoter regions. Curr Genet. 1988; 14(4):337-44. DOI: 10.1007/BF00419991. View

Hare J . Mechanisms of membrane protein turnover. Biochim Biophys Acta. 1990; 1031(1):71-90. DOI: 10.1016/0304-4157(90)90003-u. View

10.

Vida T, Huyer G, Emr S . Yeast vacuolar proenzymes are sorted in the late Golgi complex and transported to the vacuole via a prevacuolar endosome-like compartment. J Cell Biol. 1993; 121(6):1245-56. PMC: 2119698. DOI: 10.1083/jcb.121.6.1245. View

11.

Smith R, Charron M, Shah N, Lodish H, JARETT L . Immunoelectron microscopic demonstration of insulin-stimulated translocation of glucose transporters to the plasma membrane of isolated rat adipocytes and masking of the carboxyl-terminal epitope of intracellular GLUT4. Proc Natl Acad Sci U S A. 1991; 88(15):6893-7. PMC: 52195. DOI: 10.1073/pnas.88.15.6893. View

12.

Rubartelli A, Cozzolino F, Talio M, Sitia R . A novel secretory pathway for interleukin-1 beta, a protein lacking a signal sequence. EMBO J. 1990; 9(5):1503-10. PMC: 551842. DOI: 10.1002/j.1460-2075.1990.tb08268.x. View

13.

Rothblatt J, Schekman R . A hitchhiker's guide to analysis of the secretory pathway in yeast. Methods Cell Biol. 1989; 32:3-36. DOI: 10.1016/s0091-679x(08)61165-6. View

14.

Hochstrasser M, Ellison M, Chau V, Varshavsky A . The short-lived MAT alpha 2 transcriptional regulator is ubiquitinated in vivo. Proc Natl Acad Sci U S A. 1991; 88(11):4606-10. PMC: 51714. DOI: 10.1073/pnas.88.11.4606. View

15.

Seufert W, Jentsch S . Ubiquitin-conjugating enzymes UBC4 and UBC5 mediate selective degradation of short-lived and abnormal proteins. EMBO J. 1990; 9(2):543-50. PMC: 551698. DOI: 10.1002/j.1460-2075.1990.tb08141.x. View

16.

Paolini R, Kinet J . Cell surface control of the multiubiquitination and deubiquitination of high-affinity immunoglobulin E receptors. EMBO J. 1993; 12(2):779-86. PMC: 413265. DOI: 10.1002/j.1460-2075.1993.tb05712.x. View

17.

Riezman H . Yeast endocytosis. Trends Cell Biol. 1993; 3(8):273-7. DOI: 10.1016/0962-8924(93)90056-7. View

18.

Leung D, Spencer S, Cachianes G, HAMMONDS R, Collins C, Henzel W . Growth hormone receptor and serum binding protein: purification, cloning and expression. Nature. 1987; 330(6148):537-43. DOI: 10.1038/330537a0. View

19.

Davis N, Horecka J, Sprague Jr G . Cis- and trans-acting functions required for endocytosis of the yeast pheromone receptors. J Cell Biol. 1993; 122(1):53-65. PMC: 2119599. DOI: 10.1083/jcb.122.1.53. View

20.

Glotzer M, Murray A, Kirschner M . Cyclin is degraded by the ubiquitin pathway. Nature. 1991; 349(6305):132-8. DOI: 10.1038/349132a0. View