Home LiteratureArticle Details
PMID: 9151668 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Transport of a large oligomeric protein by the cytoplasm to vacuole protein targeting pathway.

The Journal of cell biology ·Vol. 137 ·No. 3 ·1997-05-05 ·Pages 609-18

Kim J, Scott SV, Oda MN, Klionsky DJ

Abstract

Aminopeptidase I (API) is transported into the yeast vacuole by the cytoplasm to vacuole targeting (Cvt) pathway. Genetic evidence suggests that autophagy, a major degradative pathway in eukaryotes, and the Cvt pathway share largely the same cellular machinery. To understand the mechanism of the Cvt import process, we examined the native state of API. Dodecameric assembly of precursor API in the cytoplasm and membrane binding were rapid events, whereas subsequent vacuolar import appeared to be rate limiting. A unique temperature-sensitive API-targeting mutant allowed us to kinetically monitor its oligomeric state during translocation. Our findings indicate that API is maintained as a dodecamer throughout its import and will be useful to study the posttranslational movement of folded proteins across biological membranes.

MeSH Terms
Aminopeptidases/metabolism Autophagy Biological Transport Cytoplasm/metabolism Fungal Proteins/metabolism Intracellular Membranes/metabolism Macromolecular Substances Protein Precursors/metabolism Protein Structure, Secondary Saccharomyces cerevisiae/enzymology,metabolism Saccharomyces cerevisiae Proteins Sequence Deletion Vacuoles/enzymology,metabolism
Chemicals
Fungal Proteins Macromolecular Substances Protein Precursors Saccharomyces cerevisiae Proteins Aminopeptidases APE1 protein, S cerevisiae
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kim J
Section of Microbiology, University of California, Davis 95616, USA.
Scott S V
Oda M N
Klionsky D J
References (46)
46 references, click to expand
  1. GTPases: multifunctional molecular switches regulating vesicular traffic.
    Annu Rev Biochem. 1994;63:949-90 PMID: 7979258
  2. The mitochondrial protein import apparatus.
    Annu Rev Biochem. 1990;59:331-53 PMID: 2197978
  3. The fungal vacuole: composition, function, and biogenesis.
    Microbiol Rev. 1990 Sep;54(3):266-92 PMID: 2215422
  4. Genetic and phenotypic overlap between autophagy and the cytoplasm to vacuole protein targeting pathway.
    J Biol Chem. 1996 Jul 26;271(30):17621-4 PMID: 8663607
  5. Protein translocation across mitochondrial membranes: what a long, strange trip it is.
    Cell. 1995 Nov 17;83(4):517-9 PMID: 7585952
  6. Yeast aminopeptidase I is post-translationally sorted from the cytosol to the vacuole by a mechanism mediated by its bipartite N-terminal extension.
    EMBO J. 1995 Nov 15;14(22):5476-84 PMID: 8521804
  7. Protein import into peroxisomes and biogenesis of the organelle.
    Annu Rev Cell Biol. 1993;9:445-78 PMID: 8280468
  8. Vacuolar/lysosomal proteolysis: proteases, substrates, mechanisms.
    Curr Opin Cell Biol. 1993 Dec;5(6):990-6 PMID: 8129953
  9. Membrane protein sorting: biosynthesis, transport and processing of yeast vacuolar alkaline phosphatase.
    EMBO J. 1989 Aug;8(8):2241-50 PMID: 2676517
  10. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis.
    Arch Biochem Biophys. 1968 Jul;126(1):155-64 PMID: 5671059
  11. Mechanisms of intralysosomal degradation with special reference to autophagocytosis and heterophagocytosis of cell organelles.
    Int Rev Cytol. 1981;73:149-82 PMID: 7028659
  12. Tracing intracellular proteolytic pathways. Proteolysis of fatty acid synthase and other cytoplasmic proteins in the yeast Saccharomyces cerevisiae.
    J Biol Chem. 1993 Dec 25;268(36):27269-76 PMID: 8262967
  13. Saccharomyces cerevisiae peroxisomal thiolase is imported as a dimer.
    Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10541-5 PMID: 7937990
  14. An oligomeric protein is imported into peroxisomes in vivo.
    J Cell Biol. 1994 Dec;127(5):1245-57 PMID: 7962087
  15. Isolation of autophagocytosis mutants of Saccharomyces cerevisiae.
    FEBS Lett. 1994 Aug 1;349(2):275-80 PMID: 8050581
  16. Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae.
    FEBS Lett. 1993 Oct 25;333(1-2):169-74 PMID: 8224160
  17. How proteins penetrate peroxisomes.
    Cell. 1995 Nov 17;83(4):525-8 PMID: 7585954
  18. Resolution of subunit interactions and cytoplasmic subcomplexes of the yeast vacuolar proton-translocating ATPase.
    J Biol Chem. 1996 Apr 26;271(17):10397-404 PMID: 8626613
  19. Reconstitution of the initial steps of mitochondrial protein import.
    Nature. 1995 Aug 24;376(6542):705-9 PMID: 7651521
  20. Guide to yeast genetics and molecular biology.
    Methods Enzymol. 1991;194:1-863 PMID: 2005781
  21. Molecular chaperones in cellular protein folding.
    Nature. 1996 Jun 13;381(6583):571-9 PMID: 8637592
  22. Protein transport across the eukaryotic endoplasmic reticulum and bacterial inner membranes.
    Annu Rev Biochem. 1996;65:271-303 PMID: 8811181
  23. Mechanism and regulation of protein degradation in liver.
    Diabetes Metab Rev. 1989 Feb;5(1):49-70 PMID: 2649336
  24. Protein sorting by transport vesicles.
    Science. 1996 Apr 12;272(5259):227-34 PMID: 8602507
  25. Transport of microinjected alcohol oxidase from Pichia pastoris into vesicles in mammalian cells: involvement of the peroxisomal targeting signal.
    J Cell Biol. 1992 Aug;118(3):499-508 PMID: 1639840
  26. The quaternary structure of yeast aminopeptidase I. 1. Molecular forms and subunit size.
    Z Naturforsch C. 1977 Nov-Dec;32(11-12):929-37 PMID: 146355
  27. Stress-induced proteolysis in yeast.
    Mol Microbiol. 1992 Sep;6(17):2437-42 PMID: 1406281
  28. Identification of a cytoplasm to vacuole targeting determinant in aminopeptidase I.
    J Cell Biol. 1996 Mar;132(6):999-1010 PMID: 8601598
  29. Autophagy and related mechanisms of lysosome-mediated protein degradation.
    Trends Cell Biol. 1994 Apr;4(4):139-43 PMID: 14731737
  30. 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
  31. Identification of tubulin from the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4962-6 PMID: 368805
  32. Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization.
    J Cell Biol. 1994 Mar;124(6):903-13 PMID: 8132712
  33. The nuclear pore complex.
    Annu Rev Biochem. 1995;64:865-96 PMID: 7574503
  34. Protein import into mitochondria: a paradigm for the translocation of polypeptides across membranes.
    Curr Opin Cell Biol. 1993 Aug;5(4):694-700 PMID: 8257609
  35. 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
  36. Cytoplasm-to-vacuole targeting and autophagy employ the same machinery to deliver proteins to the yeast vacuole.
    Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12304-8 PMID: 8901576
  37. Common principles of protein translocation across membranes.
    Science. 1996 Mar 15;271(5255):1519-26 PMID: 8599107
  38. 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
  39. In vitro reconstitution of cytoplasm to vacuole protein targeting in yeast.
    J Cell Biol. 1995 Dec;131(6 Pt 2):1727-35 PMID: 8557740
  40. Isolation and characterization of yeast mutants in the cytoplasm to vacuole protein targeting pathway.
    J Cell Biol. 1995 Nov;131(3):591-602 PMID: 7593182
  41. Nucleocytoplasmic transport.
    Science. 1996 Mar 15;271(5255):1513-8 PMID: 8599106
  42. Protein folding and association: in vitro studies for self-organization and targeting in the cell.
    Curr Top Cell Regul. 1996;34:209-314 PMID: 8646849
  43. Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction.
    J Cell Biol. 1992 Oct;119(2):301-11 PMID: 1400575
  44. Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway.
    J Cell Biol. 1992 Oct;119(2):287-99 PMID: 1400574
  45. Rab GTPases in vesicular transport.
    Curr Opin Cell Biol. 1993 Aug;5(4):613-20 PMID: 8257602
  46. Comparative studies on the dodecameric and hexameric forms of yeast aminopeptidase I.
    Z Naturforsch C. 1979 May-Jun;34C(5-6):381-6 PMID: 156998
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.137.3.609
Published
1997-05-05
Pages
609-18
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2139888
Subset
IM
Grants
NIGMS NIH HHS · GM53396 · United States
External Links
PubMed source DOI: 10.1083/jcb.137.3.609 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