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PMID: 8157009 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The Saccharomyces cerevisiae APS1 gene encodes a homolog of the small subunit of the mammalian clathrin AP-1 complex: evidence for functional interaction with clathrin at the Golgi complex.

The EMBO journal ·Vol. 13 ·No. 7 ·1994-04-01 ·Pages 1706-17

Phan HL, Finlay JA, Chu DS, Tan PK, Kirchhausen T, Payne GS

Abstract

Clathrin-associated protein (AP) complexes have been implicated in the assembly of clathrin coats and the selectivity of clathrin-mediated protein transport processes. We have identified a yeast gene, APS1, encoding a homolog of the small (referred to herein as sigma) subunits of the mammalian AP-1 complex. Sequence comparisons have shown that Aps1p is more similar to the sigma subunit of the Golgi-localized mammalian AP-1 complex than Aps2p, which is more related to the plasma membrane AP-2 sigma subunit. Like their mammalian counterparts, Aps1p and Aps2p are components of distinct, large (> 200 kDa) complexes and a significant portion of the Aps proteins co-fractionate with clathrin-coated vesicles during gel filtration chromatography. Unexpectedly, even though the evolutionary conservation of AP small subunits is substantial (50% identity between mammalian and yeast proteins), disruptions of APS1 (aps1 delta) and APS2 (aps2 delta), individually or in combination, elicit no detectable mutant phenotypes. These data indicate that the Aps proteins are not absolutely required for clathrin-mediated selective protein transport in cells expressing wild type clathrin. However, aps1 delta accentuated the slow growth and alpha-factor pheromone maturation defect of cells carrying a temperature-sensitive allele of clathrin heavy chain (Chc) (chc1-ts). In contrast, aps1 delta did not influence the effects of chc1-ts on vacuolar protein sorting or receptor-mediated endocytosis. The aps2 delta mutation resulted in a slight effect on chc1-ts cell growth but had no additional effects. The growth defect of cells completely lacking Chc was compounded by aps1 delta but not aps2 delta. These results comprise evidence that Aps1p is involved in a subset of clathrin functions at the Golgi apparatus. The effect of aps1 delta on cells devoid of clathrin function suggests that Aps1p also participates in clathrin-independent processes.

Related Genes
MeSH Terms
Adaptor Protein Complex 1 Adaptor Protein Complex alpha Subunits Adaptor Protein Complex sigma Subunits Adaptor Proteins, Vesicular Transport Amino Acid Sequence Base Sequence Biological Transport Cell Compartmentation Clathrin/metabolism Genes, Fungal/genetics Golgi Apparatus/metabolism Intracellular Membranes/chemistry Macromolecular Substances Mating Factor Molecular Sequence Data Multigene Family Peptides/metabolism Proteins/genetics,metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Deletion Sequence Homology, Amino Acid
Chemicals
APS1 protein, S cerevisiae Adaptor Protein Complex 1 Adaptor Protein Complex alpha Subunits Adaptor Protein Complex sigma Subunits Adaptor Proteins, Vesicular Transport Clathrin Macromolecular Substances Peptides Proteins Saccharomyces cerevisiae Proteins adaptor protein complex 1, sigma 1 subunit Mating Factor
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Phan H L
Molecular Biology Institute, School of Medicine, University of California, Los Angeles 90024.
Finlay J A
Chu D S
Tan P K
Kirchhausen T
Payne G S
References (65)
65 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. Clathrin: a role in the intracellular retention of a Golgi membrane protein.
    Science. 1989 Sep 22;245(4924):1358-65 PMID: 2675311
  3. Assembly polypeptides from coated vesicles mediate reassembly of unique clathrin coats.
    J Cell Biol. 1983 Nov;97(5 Pt 1):1339-47 PMID: 6138359
  4. Identification of a putative yeast homolog of the mammalian beta chains of the clathrin-associated protein complexes.
    Mol Cell Biol. 1990 Nov;10(11):6089-90 PMID: 2122239
  5. The appendage domain of the AP-2 subunit is not required for assembly or invagination of clathrin-coated pits.
    J Cell Biol. 1993 Jan;120(1):47-54 PMID: 8380176
  6. Yeast endocytosis assays.
    Methods Enzymol. 1991;194:697-710 PMID: 2005817
  7. Purification and characterization of two distinct complexes of assembly polypeptides from calf brain coated vesicles that differ in their polypeptide composition and kinase activities.
    J Biol Chem. 1987 Sep 5;262(25):12182-8 PMID: 2887566
  8. Cloning of cDNAs encoding two related 100-kD coated vesicle proteins (alpha-adaptins).
    J Cell Biol. 1989 Mar;108(3):833-42 PMID: 2564002
  9. Clathrin-coated vesicles.
    Curr Opin Cell Biol. 1989 Aug;1(4):684-90 PMID: 2576383
  10. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast.
    Methods Enzymol. 1991;194:281-301 PMID: 2005793
  11. Viability of clathrin heavy-chain-deficient Saccharomyces cerevisiae is compromised by mutations at numerous loci: implications for the suppression hypothesis.
    Mol Cell Biol. 1991 Aug;11(8):3868-78 PMID: 2072897
  12. Clathrin-coated pits contain an integral membrane protein that binds the AP-2 subunit with high affinity.
    J Biol Chem. 1990 Sep 25;265(27):16514-20 PMID: 1975814
  13. Clathrin, adaptors, and sorting.
    Annu Rev Cell Biol. 1990;6:151-71 PMID: 2177341
  14. Genetic analysis of clathrin function in yeast.
    J Membr Biol. 1990 Jun;116(2):93-105 PMID: 2199679
  15. Structural and functional division into two domains of the large (100- to 115-kDa) chains of the clathrin-associated protein complex AP-2.
    Proc Natl Acad Sci U S A. 1989 Apr;86(8):2612-6 PMID: 2495531
  16. Molecular cloning and complete amino acid sequence of AP50, an assembly protein associated with clathrin-coated vesicles.
    DNA. 1988 Dec;7(10):663-9 PMID: 3148444
  17. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets.
    Cell. 1979 Feb;16(2):303-12 PMID: 455437
  18. 100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis.
    Proc Natl Acad Sci U S A. 1989 Dec;86(23):9289-93 PMID: 2574457
  19. 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
  20. Clathrin facilitates the internalization of seven transmembrane segment receptors for mating pheromones in yeast.
    J Cell Biol. 1993 Dec;123(6 Pt 2):1707-16 PMID: 8276891
  21. Stabilization of clathrin coats by the core of the clathrin-associated protein complex AP-2.
    Biochemistry. 1990 Dec 4;29(48):10791-8 PMID: 2125494
  22. Structure and function of the yeast URA3 gene: expression in Escherichia coli.
    Gene. 1984 Jul-Aug;29(1-2):113-24 PMID: 6092217
  23. Intracellular targeting and structural conservation of a prohormone-processing endoprotease.
    Science. 1989 Oct 27;246(4929):482-6 PMID: 2683070
  24. Yeast clathrin has a distinctive light chain that is important for cell growth.
    J Cell Biol. 1990 Oct;111(4):1437-49 PMID: 2211819
  25. Co-localization of molecules involved in antigen processing and presentation in an early endocytic compartment.
    Nature. 1990 Jan 11;343(6254):133-9 PMID: 2404209
  26. Cloning and expression of gamma-adaptin, a component of clathrin-coated vesicles associated with the Golgi apparatus.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2319-26 PMID: 2126014
  27. Clathrin assembly proteins: affinity purification and a model for coat assembly.
    J Cell Biol. 1987 Nov;105(5):1989-98 PMID: 2890644
  28. A test of clathrin function in protein secretion and cell growth.
    Science. 1985 Nov 29;230(4729):1009-14 PMID: 2865811
  29. AP17 and AP19, the mammalian small chains of the clathrin-associated protein complexes show homology to Yap17p, their putative homolog in yeast.
    J Biol Chem. 1991 Jun 15;266(17):11153-7 PMID: 2040623
  30. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene.
    Gene. 1980 Jul;10(2):157-66 PMID: 6248420
  31. SEC62 encodes a putative membrane protein required for protein translocation into the yeast endoplasmic reticulum.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2653-64 PMID: 2687286
  32. Genetic and biochemical characterization of clathrin-deficient Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Nov;7(11):3888-98 PMID: 3323882
  33. Structural relationships between clathrin assembly proteins from the Golgi and the plasma membrane.
    EMBO J. 1988 Apr;7(4):919-29 PMID: 3402440
  34. Sequence of the clathrin heavy chain from Saccharomyces cerevisiae and requirement of the COOH terminus for clathrin function.
    J Cell Biol. 1991 Jan;112(1):65-80 PMID: 1898742
  35. Interaction of assembly protein AP-2 and its isolated subunits with clathrin.
    Biochemistry. 1991 Jun 4;30(22):5590-7 PMID: 2036430
  36. The nuclear migration signal of Xenopus laevis nucleoplasmin.
    EMBO J. 1987 Sep;6(9):2617-25 PMID: 3119324
  37. Adaptins.
    Trends Cell Biol. 1992 Oct;2(10):293-7 PMID: 14731924
  38. Enzymes required for yeast prohormone processing.
    Annu Rev Physiol. 1988;50:345-62 PMID: 3288097
  39. Use of T7 RNA polymerase to direct expression of cloned genes.
    Methods Enzymol. 1990;185:60-89 PMID: 2199796
  40. Cloning genes by complementation in yeast.
    Methods Enzymol. 1991;194:195-230 PMID: 2005788
  41. Selective and immediate effects of clathrin heavy chain mutations on Golgi membrane protein retention in Saccharomyces cerevisiae.
    J Cell Biol. 1992 Aug;118(3):531-40 PMID: 1322413
  42. Secretion in yeast. Purification and in vitro translocation of chemical amounts of prepro-alpha-factor.
    J Biol Chem. 1991 Jul 25;266(21):13811-4 PMID: 1856213
  43. A role for clathrin in the sorting of vacuolar proteins in the Golgi complex of yeast.
    EMBO J. 1992 Aug;11(8):2811-8 PMID: 1639056
  44. 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
  45. Determination of nucleotide sequences in DNA.
    Science. 1981 Dec 11;214(4526):1205-10 PMID: 7302589
  46. The medium chains of the mammalian clathrin-associated proteins have a homolog in yeast.
    Eur J Biochem. 1991 Dec 5;202(2):569-74 PMID: 1761056
  47. Identification of coated vesicles in Saccharomyces cerevisiae.
    J Cell Biol. 1984 Jan;98(1):341-6 PMID: 6368570
  48. Protein transport to the vacuole and receptor-mediated endocytosis by clathrin heavy chain-deficient yeast.
    J Cell Biol. 1988 May;106(5):1453-61 PMID: 2836429
  49. Characterization of the plasma membrane Mg2+-ATPase from the yeast, Saccharomyces cerevisiae.
    J Biol Chem. 1979 May 10;254(9):3326-32 PMID: 155066
  50. Yeast TATA-box transcription factor gene.
    Proc Natl Acad Sci U S A. 1989 Oct;86(20):7785-9 PMID: 2682626
  51. Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin.
    EMBO J. 1984 Sep;3(9):1951-7 PMID: 6149117
  52. Clathrin and associated assembly and disassembly proteins.
    Annu Rev Biochem. 1990;59:415-38 PMID: 1973890
  53. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  54. Conservation and diversity in families of coated vesicle adaptins.
    J Biol Chem. 1990 Mar 25;265(9):4814-20 PMID: 1969413
  55. Stage-specific assays for coated pit formation and coated vesicle budding in vitro.
    J Cell Biol. 1991 Sep;114(5):869-80 PMID: 1908470
  56. Clathrin-coated vesicle assembly polypeptides: physical properties and reconstitution studies with brain membranes.
    J Cell Biol. 1988 Jan;106(1):39-50 PMID: 2892842
  57. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments.
    Gene. 1979 Dec;8(1):17-24 PMID: 395030
  58. Reconstitution of clathrin-coated pit budding from plasma membranes.
    J Cell Biol. 1991 Sep;114(5):881-91 PMID: 1908471
  59. Deep-etch visualization of proteins involved in clathrin assembly.
    J Cell Biol. 1988 Sep;107(3):877-86 PMID: 3417785
  60. Clathrin and HA2 adaptors: effects of potassium depletion, hypertonic medium, and cytosol acidification.
    J Cell Biol. 1993 Apr;121(1):61-72 PMID: 8458873
  61. Clathrin assembly protein AP-2 induces aggregation of membrane vesicles: a possible role for AP-2 in endosome formation.
    J Cell Biol. 1992 Nov;119(4):787-96 PMID: 1358896
  62. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  63. Clathrin requirement for normal growth of yeast.
    Science. 1987 Oct 23;238(4826):504-9 PMID: 3116672
  64. Genetic analysis of the yeast cytoskeleton.
    Annu Rev Genet. 1987;21:259-84 PMID: 3327466
  65. Living with clathrin: its role in intracellular membrane traffic.
    Science. 1988 Dec 9;242(4884):1396-402 PMID: 2904698
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-04-01
Pages
1706-17
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395003
Subset
IM
Grants
NIGMS NIH HHS · GM 07185 · United States
NIGMS NIH HHS · GM 39040 · United States
NIGMS NIH HHS · GM13294 · United States
Databases
GENBANK
Z30314
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