Home LiteratureArticle Details
PMID: 8534908 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The cytoplasmic tail domain of the vacuolar protein sorting receptor Vps10p and a subset of VPS gene products regulate receptor stability, function, and localization.

Molecular biology of the cell ·Vol. 6 ·No. 9 ·1995-09-00 ·Pages 1089-102

Cereghino JL, Marcusson EG, Emr SD

Abstract

VPS10 of Saccharomyces cerevisiae encodes a type I transmembrane receptor protein required for the sorting of the soluble vacuolar hydrolase carboxypeptidase Y (CPY). To characterize the essential structural features and intercompartmental transport itinerary of the CPY receptor, we have constructed mutant forms of Vps10p that alter the carboxyterminal cytoplasmic tail of the protein. In addition, we have analyzed the effect these mutations as well as mutations in several VPS genes have on the function, stability, and localization of Vps10p. Although wild-type Vps10p is very stable over a 3-h chase period, overproduction of Vps10p results in PEP4-dependent degradation of the receptor. Immunofluorescence studies indicate that overexpressed receptor is delivered to the vacuole. A mutant form of Vps10p, in which 157 residues of the 164-residue cytoplasmic tail domain have been deleted, missorts CPY and is degraded rapidly. Additional mutations in the carboxy-terminus of Vps10p, including a deletion of a putative retention/recycling signal (FYVF), also result in CPY missorting and PEP4-dependent receptor instability. Because the cytoplasmic tail domain may interact with other factors, possibly VPS gene products, Vps10p stability was examined in a number of vps mutants. As was observed with the late Golgi protein Kex2p, Vps10p is unstable in a vps1 mutant. However, instability of Vps10p is even more severe in the class E vps mutants. Double mutant analyses demonstrate that this rapid degradation is dependent upon vacuolar proteases and a functional vacuolar ATPase. Fractionation studies of Vps10p in class E vps mutant strains indicate that the turnover of Vps10p occurs in a compartment other than the vacuole. These data are consistent with a model in which the cytoplasmic tail of Vps10p directs cycling of the receptor between a late Golgi sorting compartment and a prevacuolar endosome-like compartment, an exaggerated form of which is present in the vps class E mutants.

MeSH Terms
Carboxypeptidases/metabolism Cathepsin A Cytoplasm/metabolism Endocytosis Fungal Proteins/chemistry,genetics,metabolism Genes, Fungal Membrane Proteins/metabolism Mutagenesis, Site-Directed Protein Structure, Tertiary Receptors, Cell Surface/chemistry,genetics,metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Vacuoles/metabolism Vesicular Transport Proteins
Chemicals
Fungal Proteins Membrane Proteins PEP1 protein, S cerevisiae Receptors, Cell Surface Saccharomyces cerevisiae Proteins Vesicular Transport Proteins Carboxypeptidases Cathepsin A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Cereghino J L
Division of Cellular and Molecular Medicine, Howard Hughes Medical Institute, University of California, School of Medicine, San Diego, La Jolla 92093-0668, USA.
Marcusson E G
Emr S D
References (56)
56 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Receptor-mediated protein sorting to the vacuole in yeast: roles for a protein kinase, a lipid kinase and GTP-binding proteins.
    Annu Rev Cell Dev Biol. 1995;11:1-33 PMID: 8689553
  3. Early stages in the yeast secretory pathway are required for transport of carboxypeptidase Y to the vacuole.
    Cell. 1982 Sep;30(2):439-48 PMID: 6754086
  4. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  5. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  6. PEP4 gene function is required for expression of several vacuolar hydrolases in Saccharomyces cerevisiae.
    Genetics. 1982 Dec;102(4):665-77 PMID: 6764901
  7. The synthesis and function of proteases in Saccharomyces: genetic approaches.
    Annu Rev Genet. 1984;18:233-70 PMID: 6397123
  8. Protein sorting in yeast: mutants defective in vacuole biogenesis mislocalize vacuolar proteins into the late secretory pathway.
    Cell. 1986 Dec 26;47(6):1041-51 PMID: 3536126
  9. PEP4 gene of Saccharomyces cerevisiae encodes proteinase A, a vacuolar enzyme required for processing of vacuolar precursors.
    Mol Cell Biol. 1986 Jul;6(7):2490-9 PMID: 3023936
  10. Isolation of yeast mutants defective in protein targeting to the vacuole.
    Proc Natl Acad Sci U S A. 1986 Dec;83(23):9075-9 PMID: 3538017
  11. A ras-like protein is required for a post-Golgi event in yeast secretion.
    Cell. 1987 May 22;49(4):527-38 PMID: 3552249
  12. Intracellular sorting and processing of a yeast vacuolar hydrolase: proteinase A propeptide contains vacuolar targeting information.
    Mol Cell Biol. 1988 May;8(5):2105-16 PMID: 3290649
  13. Organelle assembly in yeast: characterization of yeast mutants defective in vacuolar biogenesis and protein sorting.
    J Cell Biol. 1988 Oct;107(4):1369-83 PMID: 3049619
  14. Protein sorting in Saccharomyces cerevisiae: isolation of mutants defective in the delivery and processing of multiple vacuolar hydrolases.
    Mol Cell Biol. 1988 Nov;8(11):4936-48 PMID: 3062374
  15. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  16. Specificity of binding of clathrin adaptors to signals on the mannose-6-phosphate/insulin-like growth factor II receptor.
    EMBO J. 1989 Apr;8(4):1041-7 PMID: 2545438
  17. Characterization of genes required for protein sorting and vacuolar function in the yeast Saccharomyces cerevisiae.
    EMBO J. 1989 Jul;8(7):2057-65 PMID: 2676511
  18. Membrane protein sorting: biosynthesis, transport and processing of yeast vacuolar alkaline phosphatase.
    EMBO J. 1989 Aug;8(8):2241-50 PMID: 2676517
  19. The fungal vacuole: composition, function, and biogenesis.
    Microbiol Rev. 1990 Sep;54(3):266-92 PMID: 2215422
  20. In vitro reconstitution of intercompartmental protein transport to the yeast vacuole.
    J Cell Biol. 1990 Dec;111(6 Pt 2):2871-84 PMID: 2269659
  21. Clathrin, adaptors, and sorting.
    Annu Rev Cell Biol. 1990;6:151-71 PMID: 2177341
  22. Localization of the signal for rapid internalization of the bovine cation-independent mannose 6-phosphate/insulin-like growth factor-II receptor to amino acids 24-29 of the cytoplasmic tail.
    J Biol Chem. 1991 Mar 25;266(9):5682-8 PMID: 1848556
  23. Immunolocalization of Kex2 protease identifies a putative late Golgi compartment in the yeast Saccharomyces cerevisiae.
    J Cell Biol. 1991 May;113(3):527-38 PMID: 2016334
  24. Compartmental organization of Golgi-specific protein modification and vacuolar protein sorting events defined in a yeast sec18 (NSF) mutant.
    J Cell Biol. 1991 Jul;114(2):207-18 PMID: 2071670
  25. Posttranslational processing of the prohormone-cleaving Kex2 protease in the Saccharomyces cerevisiae secretory pathway.
    J Cell Biol. 1991 Oct;115(2):297-307 PMID: 1918142
  26. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site.
    Anal Biochem. 1992 Jan;200(1):81-8 PMID: 1595905
  27. Structure and function of the mannose 6-phosphate/insulinlike growth factor II receptors.
    Annu Rev Biochem. 1992;61:307-30 PMID: 1323236
  28. Vesicle-mediated protein sorting.
    Annu Rev Biochem. 1992;61:471-516 PMID: 1497318
  29. Alternative pathways for the sorting of soluble vacuolar proteins in yeast: a vps35 null mutant missorts and secretes only a subset of vacuolar hydrolases.
    Mol Biol Cell. 1992 Apr;3(4):415-27 PMID: 1498362
  30. A simple and efficient procedure for transformation of yeasts.
    Biotechniques. 1992 Jul;13(1):18-20 PMID: 1503765
  31. A His-Leu-Leu sequence near the carboxyl terminus of the cytoplasmic domain of the cation-dependent mannose 6-phosphate receptor is necessary for the lysosomal enzyme sorting function.
    J Biol Chem. 1992 Aug 25;267(24):17110-5 PMID: 1324923
  32. Membrane protein sorting in the yeast secretory pathway: evidence that the vacuole may be the default compartment.
    J Cell Biol. 1992 Oct;119(1):69-83 PMID: 1527174
  33. Subunit composition, biosynthesis, and assembly of the yeast vacuolar proton-translocating ATPase.
    J Bioenerg Biomembr. 1992 Aug;24(4):383-93 PMID: 1400283
  34. Genetic and cell biological aspects of the yeast vacuolar H(+)-ATPase.
    J Bioenerg Biomembr. 1992 Aug;24(4):395-405 PMID: 1400284
  35. Structural conservation and functional diversity of V-ATPases.
    J Bioenerg Biomembr. 1992 Aug;24(4):407-14 PMID: 1400285
  36. The yeast VPS17 gene encodes a membrane-associated protein required for the sorting of soluble vacuolar hydrolases.
    J Biol Chem. 1993 Jan 5;268(1):559-69 PMID: 8416961
  37. Yeast Kex1p is a Golgi-associated membrane protein: deletions in a cytoplasmic targeting domain result in mislocalization to the vacuolar membrane.
    J Cell Biol. 1992 Dec;119(6):1459-68 PMID: 1469044
  38. Mutation of a tyrosine localization signal in the cytosolic tail of yeast Kex2 protease disrupts Golgi retention and results in default transport to the vacuole.
    Mol Biol Cell. 1992 Dec;3(12):1353-71 PMID: 1493334
  39. Morphological classification of the yeast vacuolar protein sorting mutants: evidence for a prevacuolar compartment in class E vps mutants.
    Mol Biol Cell. 1992 Dec;3(12):1389-402 PMID: 1493335
  40. The VPS16 gene product associates with a sedimentable protein complex and is essential for vacuolar protein sorting in yeast.
    J Biol Chem. 1993 Mar 5;268(7):4953-62 PMID: 8444873
  41. A membrane-associated complex containing the Vps15 protein kinase and the Vps34 PI 3-kinase is essential for protein sorting to the yeast lysosome-like vacuole.
    EMBO J. 1993 May;12(5):2195-204 PMID: 8387919
  42. Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues.
    J Cell Biol. 1993 Jun;121(6):1197-209 PMID: 8509444
  43. 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 Jun;121(6):1245-56 PMID: 8509446
  44. Vps1p, a member of the dynamin GTPase family, is necessary for Golgi membrane protein retention in Saccharomyces cerevisiae.
    EMBO J. 1993 Aug;12(8):3049-59 PMID: 8344247
  45. VPS21 encodes a rab5-like GTP binding protein that is required for the sorting of yeast vacuolar proteins.
    EMBO J. 1994 Mar 15;13(6):1297-309 PMID: 8137814
  46. Sorting of membrane proteins in the yeast secretory pathway.
    J Biol Chem. 1994 Apr 8;269(14):10185-8 PMID: 8144594
  47. The sorting receptor for yeast vacuolar carboxypeptidase Y is encoded by the VPS10 gene.
    Cell. 1994 May 20;77(4):579-86 PMID: 8187177
  48. Signal-mediated retrieval of a membrane protein from the Golgi to the ER in yeast.
    J Cell Biol. 1994 Nov;127(3):653-65 PMID: 7962050
  49. Clathrin-dependent localization of alpha 1,3 mannosyltransferase to the Golgi complex of Saccharomyces cerevisiae.
    J Cell Biol. 1994 Nov;127(3):667-78 PMID: 7962051
  50. The Saccharomyces cerevisiae MVP1 gene interacts with VPS1 and is required for vacuolar protein sorting.
    Mol Cell Biol. 1995 Mar;15(3):1671-8 PMID: 7862158
  51. Metabolic instability and constitutive endocytosis of STE6, the a-factor transporter of Saccharomyces cerevisiae.
    Mol Biol Cell. 1994 Nov;5(11):1185-98 PMID: 7865884
  52. Vacuolar sorting. Tracking down an elusive receptor.
    Curr Biol. 1994 Nov 1;4(11):1019-22 PMID: 7874484
  53. Differential effects of compartment deacidification on the targeting of membrane and soluble proteins to the vacuole in yeast.
    J Cell Sci. 1994 Oct;107 ( Pt 10):2813-24 PMID: 7876349
  54. Endocytosis. Ringing necks with dynamin.
    Nature. 1995 Mar 9;374(6518):116-7 PMID: 7877678
  55. Golgi and vacuolar membrane proteins reach the vacuole in vps1 mutant yeast cells via the plasma membrane.
    J Cell Biol. 1995 Apr;129(1):35-46 PMID: 7698993
  56. Proteinase mutants of Saccharomyces cerevisiae.
    Genetics. 1977 Jan;85(1):23-33 PMID: 320092
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.6.9.1089
Published
1995-09-00
Pages
1089-102
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC301269
Subset
IM
Grants
NIGMS NIH HHS · GM15638 · United States
NIGMS NIH HHS · GM32703 · United States
External Links
PubMed source DOI: 10.1091/mbc.6.9.1089 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2