Home LiteratureArticle Details
PMID: 1498362 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Alternative pathways for the sorting of soluble vacuolar proteins in yeast: a vps35 null mutant missorts and secretes only a subset of vacuolar hydrolases.

Molecular biology of the cell ·Vol. 3 ·No. 4 ·1992-04-00 ·Pages 415-27

Paravicini G, Horazdovsky BF, Emr SD

Abstract

vps35 mutants of Saccharomyces cerevisiae exhibit severe defects in the localization of carboxypeptidase Y, a soluble vacuolar hydrolase. We have cloned the wild-type VPS35 gene by complementation of the vacuolar protein sorting defect exhibited by the vps35-17 mutant. Sequence analysis revealed an open reading frame predicted to encode a protein of 937 amino acids that lacks any obvious hydrophobic domains. Subcellular fractionation studies indicated that 80% of Vps35p peripherally associates with a membranous particulate cell fraction. The association of Vps35p with this fraction appears to be saturable; when overproduced, the vast majority of Vps35p remains in a soluble fraction. Disruption of the VPS35 gene demonstrated that it is not essential for yeast cell growth. However, the null allele of VPS35 results in a differential defect in the sorting of vacuolar carboxypeptidase Y (CPY), proteinase A (PrA), proteinase B (PrB), and alkaline phosphatase (ALP). proCPY was quantitatively missorted and secreted by delta vps35 cells, whereas almost all of proPrA, proPrB, and proALP were retained within the cell and converted to their mature forms, indicating delivery to the vacuole. Based on these observations, we propose that alternative pathways exist for the sorting and/or delivery of proteins to the vacuole.

MeSH Terms
Alkaline Phosphatase/biosynthesis Amino Acid Sequence Aspartic Acid Endopeptidases/biosynthesis Base Sequence Carboxypeptidases/biosynthesis Carrier Proteins Cathepsin A Fungal Proteins/genetics,metabolism Hydrolases/biosynthesis Intracellular Membranes/metabolism Membrane Proteins/genetics,metabolism Molecular Sequence Data Protein Precursors/metabolism Protein Processing, Post-Translational Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Serine Endopeptidases/biosynthesis Subcellular Fractions/chemistry Vacuoles/metabolism Vesicular Transport Proteins
Chemicals
Carrier Proteins Fungal Proteins Membrane Proteins Protein Precursors Saccharomyces cerevisiae Proteins VPS35 protein, S cerevisiae Vesicular Transport Proteins Hydrolases Alkaline Phosphatase Carboxypeptidases Cathepsin A PRC1 protein, S cerevisiae serine carboxypeptidase Serine Endopeptidases yeast proteinase B aspartic proteinase A PEP4 protein, S cerevisiae Aspartic Acid Endopeptidases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Paravicini G
Division of Biology, California Institute of Technology, Pasadena 91125.
Horazdovsky B F
Emr S D
References (54)
54 references, click to expand
  1. Nucleolar and nuclear envelope proteins of the yeast Saccharomyces cerevisiae.
    Eur J Cell Biol. 1988 Aug;46(3):554-63 PMID: 3053175
  2. Protein sorting in yeast: the localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide.
    Cell. 1987 Mar 13;48(5):887-97 PMID: 3028649
  3. Proteinase mutants of Saccharomyces cerevisiae.
    Genetics. 1977 Jan;85(1):23-33 PMID: 320092
  4. A novel protein kinase homolog essential for protein sorting to the yeast lysosome-like vacuole.
    Cell. 1991 Jan 25;64(2):425-37 PMID: 1988155
  5. A putative GTP binding protein homologous to interferon-inducible Mx proteins performs an essential function in yeast protein sorting.
    Cell. 1990 Jun 15;61(6):1063-74 PMID: 2112425
  6. The fungal vacuole: composition, function, and biogenesis.
    Microbiol Rev. 1990 Sep;54(3):266-92 PMID: 2215422
  7. In vitro reconstitution of intercompartmental protein transport to the yeast vacuole.
    J Cell Biol. 1990 Dec;111(6 Pt 2):2871-84 PMID: 2269659
  8. The biogenesis of lysosomes.
    Annu Rev Cell Biol. 1989;5:483-525 PMID: 2557062
  9. Fractionation of yeast organelles.
    Methods Cell Biol. 1989;31:335-56 PMID: 2674627
  10. Membrane protein sorting: biosynthesis, transport and processing of yeast vacuolar alkaline phosphatase.
    EMBO J. 1989 Aug;8(8):2241-50 PMID: 2676517
  11. Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast.
    Cell. 1987 May 8;49(3):295-7 PMID: 2882858
  12. Rapid and sensitive protein similarity searches.
    Science. 1985 Mar 22;227(4693):1435-41 PMID: 2983426
  13. Trafficking of lysosomal enzymes in normal and disease states.
    J Clin Invest. 1986 Jan;77(1):1-6 PMID: 3003148
  14. 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
  15. 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
  16. 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
  17. 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
  18. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
    Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12 PMID: 4504350
  19. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine.
    Science. 1981 Dec 4;214(4525):1125-9 PMID: 6272395
  20. DNA sequence required for efficient transcription termination in yeast.
    Cell. 1982 Mar;28(3):563-73 PMID: 6280875
  21. A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast.
    Cell. 1988 Jun 3;53(5):753-68 PMID: 3131018
  22. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  23. Overproduction-induced mislocalization of a yeast vacuolar protein allows isolation of its structural gene.
    Proc Natl Acad Sci U S A. 1986 May;83(10):3248-52 PMID: 3517855
  24. 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
  25. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  26. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  27. Expression of the Herpes simplex virus thymidine kinase gene in Saccharomyces cerevisiae.
    Mol Gen Genet. 1981;184(3):386-93 PMID: 6278252
  28. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast.
    Methods Enzymol. 1983;100:293-308 PMID: 6312261
  29. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  30. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  31. Asparagine-linked carbohydrate does not determine the cellular location of yeast vacuolar nonspecific alkaline phosphatase.
    J Bacteriol. 1982 Nov;152(2):865-73 PMID: 6813317
  32. Carbohydrate-free carboxypeptidase Y is transferred into the lysosome-like yeast vacuole.
    Biochem Biophys Res Commun. 1982 Feb 11;104(3):950-6 PMID: 7041906
  33. A Critical Evaluation of the Nitrogen Assimilation Tests Commonly Used in the Classification of Yeasts.
    J Bacteriol. 1946 Sep;52(3):293-301 PMID: 16561177
  34. Glycosylation and processing of prepro-alpha-factor through the yeast secretory pathway.
    Cell. 1984 Feb;36(2):309-18 PMID: 6420074
  35. 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
  36. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.
    J Mol Biol. 1980 Apr;138(2):179-207 PMID: 6997493
  37. Beta-D-fructofuranoside fructohydrolase from yeast.
    Methods Enzymol. 1975;42:504-11 PMID: 237205
  38. Phosphohexosyl components of a lysosomal enzyme are recognized by pinocytosis receptors on human fibroblasts.
    Proc Natl Acad Sci U S A. 1977 May;74(5):2026-30 PMID: 266721
  39. Identification of procollagen mRNAs transferred to diazobenzyloxymethyl paper from formaldehyde agarose gels.
    Nucleic Acids Res. 1979 Aug 10;6(11):3559-67 PMID: 573889
  40. A genetic and structural analysis of the yeast Vps15 protein kinase: evidence for a direct role of Vps15p in vacuolar protein delivery.
    EMBO J. 1991 Dec;10(13):4049-60 PMID: 1756716
  41. 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
  42. A new class of lysosomal/vacuolar protein sorting signals.
    J Biol Chem. 1990 Apr 5;265(10):5349-52 PMID: 2180925
  43. Characterization of yeast Vps33p, a protein required for vacuolar protein sorting and vacuole biogenesis.
    Mol Cell Biol. 1990 Sep;10(9):4638-49 PMID: 2201898
  44. Characterization of VPS34, a gene required for vacuolar protein sorting and vacuole segregation in Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 Dec;10(12):6742-54 PMID: 2247081
  45. Fluorescence microscopy methods for yeast.
    Methods Cell Biol. 1989;31:357-435 PMID: 2476649
  46. Biosynthesis of inositol in yeast. Primary structure of myo-inositol-1-phosphate synthase (EC 5.5.1.4) and functional analysis of its structural gene, the INO1 locus.
    J Biol Chem. 1989 Jan 15;264(2):1274-83 PMID: 2642902
  47. Processing pathway for protease B of Saccharomyces cerevisiae.
    J Cell Biol. 1989 Feb;108(2):309-25 PMID: 2645294
  48. 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
  49. Genetic map of Saccharomyces cerevisiae, edition 10.
    Yeast. 1989 Sep-Oct;5(5):321-403 PMID: 2678811
  50. Functional compartments of the yeast Golgi apparatus are defined by the sec7 mutation.
    EMBO J. 1989 Sep;8(9):2695-702 PMID: 2684655
  51. Assembly of the mitochondrial membrane system. CBP6, a yeast nuclear gene necessary for synthesis of cytochrome b.
    J Biol Chem. 1985 Feb 10;260(3):1513-20 PMID: 2981859
  52. Arrangement of genes TRP1 and TRP3 of Saccharomyces cerevisiae strains.
    Arch Microbiol. 1985 Sep;142(4):383-8 PMID: 2998296
  53. Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease.
    Cell. 1987 Mar 13;48(5):875-85 PMID: 3028648
  54. Yeast cytochrome c messenger RNA. In vitro translation and specific immunoprecipitation of the CYC1 gene product.
    J Biol Chem. 1976 Oct 25;251(20):6320-6 PMID: 185210
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.3.4.415
Published
1992-04-00
Pages
415-27
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC275592
Subset
IM
Grants
NIGMS NIH HHS · GM-12848 · United States
NIGMS NIH HHS · GM-32703 · United States
External Links
PubMed source DOI: 10.1091/mbc.3.4.415 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