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Mol Biol Cell. 1995 January; 6(1): 41–58.
PMCID: PMC275813

A late Golgi sorting function for Saccharomyces cerevisiae Apm1p, but not for Apm2p, a second yeast clathrin AP medium chain-related protein.


Mammalian clathrin-associated protein (AP) complexes, AP-1 (trans-Golgi network) and AP-2 (plasma membrane), are composed of two large subunits of 91-107 kDa, one medium chain (mu) of 47-50 kDa and one small chain (sigma) of 17-19 kDa. Two yeast genes, APM1 and APM2, have been identified that encode proteins related to AP mu chains. APM1, whose sequence was reported previously, codes for a protein of 54 kDa that has greatest similarity to the mammalian 47-kDa mu 1 chain of AP-1. APM2 encodes an AP medium chain-related protein of 605 amino acids (predicted molecular weight of 70 kDa) that is only 30-33% identical to the other family members. In yeast containing a normal clathrin heavy chain gene (CHC1), disruptions of the APM genes, singly or in combination, had no detectable phenotypic consequences. However, deletion of APM1 greatly enhanced the temperature-sensitive growth phenotype and the alpha-factor processing defect displayed by cells carrying a temperature-sensitive allele of the clathrin heavy chain gene. In contrast, deletion of APM2 caused no synthetic phenotypes with clathrin mutants. Biochemical analysis indicated that Apm1p and Apm2p are components of distinct high molecular weight complexes. Apm1p, Apm2p, and clathrin cofractionated in a discrete vesicle population, and the association of Apm1p with the vesicles was disrupted in CHC1 deletion strains. These results suggest that Apm1p is a component of an AP-1-like complex that participates with clathrin in sorting at the trans-Golgi in yeast. We propose that Apm2p represents a new class of AP-medium chain-related proteins that may be involved in a nonclathrin-mediated vesicular transport process in eukaryotic cells.

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  • Ahle S, Ungewickell E. Identification of a clathrin binding subunit in the HA2 adaptor protein complex. J Biol Chem. 1989 Nov 25;264(33):20089–20093. [PubMed]
  • Ahle S, Mann A, Eichelsbacher U, Ungewickell E. Structural relationships between clathrin assembly proteins from the Golgi and the plasma membrane. EMBO J. 1988 Apr;7(4):919–929. [PubMed]
  • Atcheson CL, DiDomenico B, Frackman S, Esposito RE, Elder RT. Isolation, DNA sequence, and regulation of a meiosis-specific eukaryotic recombination gene. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8035–8039. [PubMed]
  • Barlowe C, Orci L, Yeung T, Hosobuchi M, Hamamoto S, Salama N, Rexach MF, Ravazzola M, Amherdt M, Schekman R. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell. 1994 Jun 17;77(6):895–907. [PubMed]
  • Beltzer JP, Spiess M. In vitro binding of the asialoglycoprotein receptor to the beta adaptin of plasma membrane coated vesicles. EMBO J. 1991 Dec;10(12):3735–3742. [PubMed]
  • Chang MP, Mallet WG, Mostov KE, Brodsky FM. Adaptor self-aggregation, adaptor-receptor recognition and binding of alpha-adaptin subunits to the plasma membrane contribute to recruitment of adaptor (AP2) components of clathrin-coated pits. EMBO J. 1993 May;12(5):2169–2180. [PubMed]
  • Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. [PubMed]
  • Cryer DR, Eccleshall R, Marmur J. Isolation of yeast DNA. Methods Cell Biol. 1975;12:39–44. [PubMed]
  • Daignan-Fornier B, Bolotin-Fukuhara M. Functional exploration of the yeast (Saccharomyces cerevisiae) genome: use of a mini-Mu transposon to analyse randomly cloned DNA sequences. Yeast. 1989 Jul-Aug;5(4):259–269. [PubMed]
  • Duden R, Griffiths G, Frank R, Argos P, Kreis TE. Beta-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to beta-adaptin. Cell. 1991 Feb 8;64(3):649–665. [PubMed]
  • Gallusser A, Kirchhausen T. The beta 1 and beta 2 subunits of the AP complexes are the clathrin coat assembly components. EMBO J. 1993 Dec 15;12(13):5237–5244. [PubMed]
  • Gergen JP, Stern RH, Wensink PC. Filter replicas and permanent collections of recombinant DNA plasmids. Nucleic Acids Res. 1979 Dec 20;7(8):2115–2136. [PMC free article] [PubMed]
  • Glickman JN, Conibear E, Pearse BM. 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):1041–1047. [PubMed]
  • Heuser JE, Keen J. Deep-etch visualization of proteins involved in clathrin assembly. J Cell Biol. 1988 Sep;107(3):877–886. [PMC free article] [PubMed]
  • Heuser J, Kirchhausen T. Deep-etch views of clathrin assemblies. J Ultrastruct Res. 1985 Jul-Aug;92(1-2):1–27. [PubMed]
  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A. Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast. 1986 Sep;2(3):163–167. [PubMed]
  • Hosobuchi M, Kreis T, Schekman R. SEC21 is a gene required for ER to Golgi protein transport that encodes a subunit of a yeast coatomer. Nature. 1992 Dec 10;360(6404):603–605. [PubMed]
  • Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. [PMC free article] [PubMed]
  • Keen JH. Clathrin assembly proteins: affinity purification and a model for coat assembly. J Cell Biol. 1987 Nov;105(5):1989–1998. [PMC free article] [PubMed]
  • Keen JH. Clathrin and associated assembly and disassembly proteins. Annu Rev Biochem. 1990;59:415–438. [PubMed]
  • Keen JH, Willingham MC, Pastan IH. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell. 1979 Feb;16(2):303–312. [PubMed]
  • Kirchhausen T. Identification of a putative yeast homolog of the mammalian beta chains of the clathrin-associated protein complexes. Mol Cell Biol. 1990 Nov;10(11):6089–6090. [PMC free article] [PubMed]
  • Kirchhausen T, Nathanson KL, Matsui W, Vaisberg A, Chow EP, Burne C, Keen JH, Davis AE. 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):2612–2616. [PubMed]
  • Kirchhausen T, Davis AC, Frucht S, Greco BO, Payne GS, Tubb B. 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):11153–11157. [PubMed]
  • Koerner TJ, Hill JE, Myers AM, Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. [PubMed]
  • Kuge O, Hara-Kuge S, Orci L, Ravazzola M, Amherdt M, Tanigawa G, Wieland FT, Rothman JE. zeta-COP, a subunit of coatomer, is required for COP-coated vesicle assembly. J Cell Biol. 1993 Dec;123(6 Pt 2):1727–1734. [PMC free article] [PubMed]
  • Le Borgne R, Schmidt A, Mauxion F, Griffiths G, Hoflack B. Binding of AP-1 Golgi adaptors to membranes requires phosphorylated cytoplasmic domains of the mannose 6-phosphate/insulin-like growth factor II receptor. J Biol Chem. 1993 Oct 25;268(30):22552–22556. [PubMed]
  • Lee J, Jongeward GD, Sternberg PW. unc-101, a gene required for many aspects of Caenorhabditis elegans development and behavior, encodes a clathrin-associated protein. Genes Dev. 1994 Jan;8(1):60–73. [PubMed]
  • Lemmon SK, Jones EW. Clathrin requirement for normal growth of yeast. Science. 1987 Oct 23;238(4826):504–509. [PubMed]
  • Lemmon S, Lemmon VP, Jones EW. Characterization of yeast clathrin and anticlathrin heavy-chain monoclonal antibodies. J Cell Biochem. 1988 Apr;36(4):329–340. [PubMed]
  • Lemmon SK, Freund C, Conley K, Jones EW. Genetic instability of clathrin-deficient strains of Saccharomyces cerevisiae. Genetics. 1990 Jan;124(1):27–38. [PubMed]
  • Lemmon SK, Pellicena-Palle A, Conley K, Freund CL. 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. [PMC free article] [PubMed]
  • Mahaffey DT, Peeler JS, Brodsky FM, Anderson RG. 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):16514–16520. [PubMed]
  • Manfredi JJ, Bazari WL. 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):12182–12188. [PubMed]
  • Matsui W, Kirchhausen T. Stabilization of clathrin coats by the core of the clathrin-associated protein complex AP-2. Biochemistry. 1990 Dec 4;29(48):10791–10798. [PubMed]
  • Mortimer RK, Johnston JR. Genealogy of principal strains of the yeast genetic stock center. Genetics. 1986 May;113(1):35–43. [PubMed]
  • Mortimer RK, Contopoulou CR, King JS. Genetic and physical maps of Saccharomyces cerevisiae, Edition 11. Yeast. 1992 Oct;8(10):817–902. [PubMed]
  • Mueller SC, Branton D. Identification of coated vesicles in Saccharomyces cerevisiae. J Cell Biol. 1984 Jan;98(1):341–346. [PMC free article] [PubMed]
  • Munn AL, Silveira L, Elgort M, Payne GS. 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):3868–3878. [PMC free article] [PubMed]
  • Nakai M, Takada T, Endo T. Cloning of the YAP19 gene encoding a putative yeast homolog of AP19, the mammalian small chain of the clathrin-assembly proteins. Biochim Biophys Acta. 1993 Sep 23;1174(3):282–284. [PubMed]
  • Nakayama Y, Goebl M, O'Brine Greco B, Lemmon S, Pingchang Chow E, Kirchhausen T. The medium chains of the mammalian clathrin-associated proteins have a homolog in yeast. Eur J Biochem. 1991 Dec 5;202(2):569–574. [PubMed]
  • Nelson KK, Lemmon SK. Suppressors of clathrin deficiency: overexpression of ubiquitin rescues lethal strains of clathrin-deficient Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jan;13(1):521–532. [PMC free article] [PubMed]
  • Payne GS, Baker D, van Tuinen E, Schekman R. Protein transport to the vacuole and receptor-mediated endocytosis by clathrin heavy chain-deficient yeast. J Cell Biol. 1988 May;106(5):1453–1461. [PMC free article] [PubMed]
  • Payne GS, Schekman R. A test of clathrin function in protein secretion and cell growth. Science. 1985 Nov 29;230(4729):1009–1014. [PubMed]
  • Payne GS, Schekman R. Clathrin: a role in the intracellular retention of a Golgi membrane protein. Science. 1989 Sep 22;245(4924):1358–1365. [PubMed]
  • Pearse BM. Receptors compete for adaptors found in plasma membrane coated pits. EMBO J. 1988 Nov;7(11):3331–3336. [PubMed]
  • Pearse BM, Robinson MS. Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin. EMBO J. 1984 Sep;3(9):1951–1957. [PubMed]
  • Pearse BM, Robinson MS. Clathrin, adaptors, and sorting. Annu Rev Cell Biol. 1990;6:151–171. [PubMed]
  • Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. [PubMed]
  • Peeler JS, Donzell WC, Anderson RG. 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. [PMC free article] [PubMed]
  • Pevsner J, Volknandt W, Wong BR, Scheller RH. Two rat homologs of clathrin-associated adaptor proteins. Gene. 1994 Sep 2;146(2):279–283. [PubMed]
  • Phan HL, Finlay JA, Chu DS, Tan PK, Kirchhausen T, Payne GS. 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. EMBO J. 1994 Apr 1;13(7):1706–1717. [PubMed]
  • Prasad K, Keen JH. Interaction of assembly protein AP-2 and its isolated subunits with clathrin. Biochemistry. 1991 Jun 4;30(22):5590–5597. [PubMed]
  • Roberts CJ, Nothwehr SF, Stevens TH. 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. [PMC free article] [PubMed]
  • Robinson MS. 100-kD coated vesicle proteins: molecular heterogeneity and intracellular distribution studied with monoclonal antibodies. J Cell Biol. 1987 Apr;104(4):887–895. [PMC free article] [PubMed]
  • Robinson MS. Cloning of cDNAs encoding two related 100-kD coated vesicle proteins (alpha-adaptins). J Cell Biol. 1989 Mar;108(3):833–842. [PMC free article] [PubMed]
  • Robinson MS. Assembly and targeting of adaptin chimeras in transfected cells. J Cell Biol. 1993 Oct;123(1):67–77. [PMC free article] [PubMed]
  • Robinson MS, Kreis TE. Recruitment of coat proteins onto Golgi membranes in intact and permeabilized cells: effects of brefeldin A and G protein activators. Cell. 1992 Apr 3;69(1):129–138. [PubMed]
  • Rose MD, Broach JR. Cloning genes by complementation in yeast. Methods Enzymol. 1991;194:195–230. [PubMed]
  • Rose MD, Novick P, Thomas JH, Botstein D, Fink GR. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. [PubMed]
  • Rothman JE, Orci L. Molecular dissection of the secretory pathway. Nature. 1992 Jan 30;355(6359):409–415. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Schmid SL. The mechanism of receptor-mediated endocytosis: more questions than answers. Bioessays. 1992 Sep;14(9):589–596. [PubMed]
  • Schröder S, Ungewickell E. Subunit interaction and function of clathrin-coated vesicle adaptors from the Golgi and the plasma membrane. J Biol Chem. 1991 Apr 25;266(12):7910–7918. [PubMed]
  • Seaman MN, Ball CL, Robinson MS. Targeting and mistargeting of plasma membrane adaptors in vitro. J Cell Biol. 1993 Dec;123(5):1093–1105. [PMC free article] [PubMed]
  • Seeger M, Payne GS. A role for clathrin in the sorting of vacuolar proteins in the Golgi complex of yeast. EMBO J. 1992 Aug;11(8):2811–2818. [PubMed]
  • Seeger M, Payne GS. Selective and immediate effects of clathrin heavy chain mutations on Golgi membrane protein retention in Saccharomyces cerevisiae. J Cell Biol. 1992 Aug;118(3):531–540. [PMC free article] [PubMed]
  • Serafini T, Stenbeck G, Brecht A, Lottspeich F, Orci L, Rothman JE, Wieland FT. A coat subunit of Golgi-derived non-clathrin-coated vesicles with homology to the clathrin-coated vesicle coat protein beta-adaptin. Nature. 1991 Jan 17;349(6306):215–220. [PubMed]
  • Sikorski RS, Hieter P. 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. [PubMed]
  • Silveira LA, Wong DH, Masiarz FR, Schekman R. Yeast clathrin has a distinctive light chain that is important for cell growth. J Cell Biol. 1990 Oct;111(4):1437–1449. [PMC free article] [PubMed]
  • Smythe E, Carter LL, Schmid SL. Cytosol- and clathrin-dependent stimulation of endocytosis in vitro by purified adaptors. J Cell Biol. 1992 Dec;119(5):1163–1171. [PMC free article] [PubMed]
  • Sorkin A, Carpenter G. Interaction of activated EGF receptors with coated pit adaptins. Science. 1993 Jul 30;261(5121):612–615. [PubMed]
  • Sosa MA, Schmidt B, von Figura K, Hille-Rehfeld A. In vitro binding of plasma membrane-coated vesicle adaptors to the cytoplasmic domain of lysosomal acid phosphatase. J Biol Chem. 1993 Jun 15;268(17):12537–12543. [PubMed]
  • Stamnes MA, Rothman JE. The binding of AP-1 clathrin adaptor particles to Golgi membranes requires ADP-ribosylation factor, a small GTP-binding protein. Cell. 1993 Jun 4;73(5):999–1005. [PubMed]
  • Traub LM, Ostrom JA, Kornfeld S. Biochemical dissection of AP-1 recruitment onto Golgi membranes. J Cell Biol. 1993 Nov;123(3):561–573. [PMC free article] [PubMed]
  • Unanue ER, Ungewickell E, Branton D. The binding of clathrin triskelions to membranes from coated vesicles. Cell. 1981 Nov;26(3 Pt 1):439–446. [PubMed]
  • Vigers GP, Crowther RA, Pearse BM. Location of the 100 kd-50 kd accessory proteins in clathrin coats. EMBO J. 1986 Sep;5(9):2079–2085. [PubMed]
  • Virshup DM, Bennett V. Clathrin-coated vesicle assembly polypeptides: physical properties and reconstitution studies with brain membranes. J Cell Biol. 1988 Jan;106(1):39–50. [PMC free article] [PubMed]
  • Wang LH, Rothberg KG, Anderson RG. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol. 1993 Dec;123(5):1107–1117. [PMC free article] [PubMed]
  • Waters MG, Serafini T, Rothman JE. 'Coatomer': a cytosolic protein complex containing subunits of non-clathrin-coated Golgi transport vesicles. Nature. 1991 Jan 17;349(6306):248–251. [PubMed]
  • White MJ, Hirsch JP, Henry SA. The OPI1 gene of Saccharomyces cerevisiae, a negative regulator of phospholipid biosynthesis, encodes a protein containing polyglutamine tracts and a leucine zipper. J Biol Chem. 1991 Jan 15;266(2):863–872. [PubMed]
  • Wilcox CA, Redding K, Wright R, Fuller RS. 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):1353–1371. [PMC free article] [PubMed]
  • Wong DH, Brodsky FM. 100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties. J Cell Biol. 1992 Jun;117(6):1171–1179. [PMC free article] [PubMed]
  • Zaremba S, Keen JH. Assembly polypeptides from coated vesicles mediate reassembly of unique clathrin coats. J Cell Biol. 1983 Nov;97(5 Pt 1):1339–1347. [PMC free article] [PubMed]

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