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Cited by 320Cellular transport and secretionCaveolin-1 and cellular processesAutophagy in Disease and Therapy

Clathrin Assembly Lymphoid Myeloid Leukemia (CALM) Protein: Localization in Endocytic-coated Pits, Interactions with Clathrin, and the Impact of Overexpression on Clathrin-mediated Traffic

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Tebar, Francesc, Bohlander, Stefan K., Sorkin, Alexander·1999·Molecular Biology of the Cell
DOI 10.1091/mbc.10.8.2687PMC PMC25500
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Abstract

The clathrin assembly lymphoid myeloid leukemia (CALM) gene encodes a putative homologue of the clathrin assembly synaptic protein AP180. Hence the biochemical properties, the subcellular localization, and the role in endocytosis of a CALM protein were studied. In vitro binding and coimmunoprecipitation demonstrated that the clathrin heavy chain is the major binding partner of CALM. The bulk of cellular CALM was associated with the membrane fractions of the cell and localized to clathrin-coated areas of the plasma membrane. In the membrane fraction, CALM was present at near stoichiometric amounts relative to clathrin. To perform structure–function analysis of CALM, we engineered chimeric fusion proteins of CALM and its fragments with the green fluorescent protein (GFP). GFP–CALM was targeted to the plasma membrane–coated pits and also found colocalized with clathrin in the Golgi area. High levels of expression of GFP–CALM or its fragments with clathrin-binding activity inhibited the endocytosis of transferrin and epidermal growth factor receptors and altered the steady-state distribution of the mannose-6-phosphate receptor in the cell. In addition, GFP–CALM overexpression caused the loss of clathrin accumulation in the trans-Golgi network area, whereas the localization of the clathrin adaptor protein complex 1 in the trans-Golgi network remained unaffected. The ability of the GFP-tagged fragments of CALM to affect clathrin-mediated processes correlated with the targeting of the fragments to clathrin-coated areas and their clathrin-binding capacities. Clathrin–CALM interaction seems to be regulated by multiple contact interfaces. The C-terminal part of CALM binds clathrin heavy chain, although the full-length protein exhibited maximal ability for interaction. Altogether, the data suggest that CALM is an important component of coated pit internalization machinery, possibly involved in the regulation of clathrin recruitment to the membrane and/or the formation of the coated pit.

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References 37 · Cited by 320

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  1. Ahle S, Ungewickell E. Purification and properties of a new clathrin assembly protein. EMBO J. 1986;5:3143–3149. doi: 10.1002/j.1460-2075.1986.tb04621.x. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  2. Brodsky FM. Clathrin structure characterized with monoclonal antibodies. I. Analysis of multiple antigenic sites. J Cell Biol. 1985;101:2047–2054. doi: 10.1083/jcb.101.6.2047. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  3. Carpenter G, Cohen S. 125I-Labeled human epidermal growth factor: binding internalization, and degradation in human fibroblasts. J Cell Biol. 1976;71:159–171. doi: 10.1083/jcb.71.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  4. Chen H, Fre S, Slepnev VI, Capua MR, Takei K, Butler MH, Di Fiore PP, De Camilli P. Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis. Nature. 1998;394:793–797. doi: 10.1038/29555. [DOI] [PubMed] [Google Scholar] · DOI
  5. De Beer T, Carter RE, Lobel-Rice KE, Sorkin A, Overduin M. Structure and Asn-Pro-Phe binding pocket of the eps15 homology domain. Science. 1998;281:1357–1360. doi: 10.1126/science.281.5381.1357. [DOI] [PubMed] [Google Scholar] · DOI
  6. Dell’Angelica EC, Klumperman J, Stoorvogel W, Bonifacino JS. Association of the AP-3 adaptor complex with clathrin. Science. 1998;280:431–434. doi: 10.1126/science.280.5362.431. [DOI] [PubMed] [Google Scholar] · DOI
  7. Dreyling MH, Martinez-Climent JA, Zheng M, Mao J, Rowley JD, Bohlander SK. The t(10,11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family. Proc Natl Acad Sci USA. 1996;93:4804–4809. doi: 10.1073/pnas.93.10.4804. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI · Paper page
  8. Gill SC, von Hippel PH. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989;182:319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar] · DOI
  9. Goodman OB, Jr, Krupnick JG, Gurevich VV, Benovic JL, Keen JH. Arrestin/clathrin interaction. Localization of the arrestin binding locus to the clathrin terminal domain. J Biol Chem. 1997;272:15017–15022. doi: 10.1074/jbc.272.23.15017. [DOI] [PubMed] [Google Scholar] · DOI
  10. Goud B, Huet C, Louvard D. Assembled and unassembled pools of clathrin: a quantitative study using an enzyme immunoassay. J Cell Biol. 1985;100:521–527. doi: 10.1083/jcb.100.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  11. Hao W, Tan Z, Prasad K, Reddy KK, Chen J, Prestwich GD, Falck JR, Shears SB, Lafer EM. Regulation of AP-3 function by inositides. Identification of phosphatidylinositol 3,4,5-triphosphate as a potent ligand. J Biol Chem. 1997;272:6393–6398. doi: 10.1074/jbc.272.10.6393. [DOI] [PubMed] [Google Scholar] · DOI
  12. Keen JH, Black MM. The phosphorylation of coated membrane proteins in intact neurons. J Cell Biol. 1986;102:1325–1333. doi: 10.1083/jcb.102.4.1325. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  13. Kohtz DS, Puszkin S. A neuronal protein (NP185) associated with clathrin-coated vesicles. Characterization of NP185 with monoclonal antibodies. J Biol Chem. 1988;263:7418–7425. [PubMed] [Google Scholar]
  14. Lindner R, Ungewickell E. Clathrin-associated proteins of bovine brain coated vesicles. An analysis of their number and assembly-promoting activity. J Biol Chem. 1992;267:16567–16573. [PubMed] [Google Scholar]
  15. Murphy JE, Keen JH. Recognition sites for clathrin-associated proteins AP-2 and AP-3 on clathrin triskelia. J Biol Chem. 1992;267:10850–10855. [PubMed] [Google Scholar]
  16. Murphy JE, Pleasure IT, Puszkin S, Prasad K, Keen JH. Clathrin assembly protein AP-3. The identity of the 155K protein, AP180, and NP185 and demonstration of a clathrin binding domain. J Biol Chem. 1991;266:4401–4408. [PubMed] [Google Scholar]
  17. Norris FA, Ungewickell E, Majerus PW. Inositol hexakisphosphate binds to clathrin assembly protein 3(AP-3/AP180) and inhibits clathrin cage assembly in vitro. J Biol Chem. 1995;270:214–217. doi: 10.1074/jbc.270.1.214. [DOI] [PubMed] [Google Scholar] · DOI
  18. Odorizzi G, Cowles CR, Emr SD. The AP-3 complex: a coat of many colors. Trends Cell Biol. 1998;8:282–288. doi: 10.1016/s0962-8924(98)01295-1. [DOI] [PubMed] [Google Scholar] · DOI
  19. Robinson MS. Coats and vesicle budding. Trends Cell Biol. 1997;7:99–102. doi: 10.1016/S0962-8924(96)10048-9. [DOI] [PubMed] [Google Scholar] · DOI
  20. Salcini AE, Confalonieri S, Doria M, Santolini E, Tassi E, Minenkova O, Cesareni G, Pelicci PG, Di Fiore PP. Binding specificity and in vivo targets of the EH domain, a novel protein-protein interaction module. Genes Dev. 1997;11:2239–2249. doi: 10.1101/gad.11.17.2239. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI · Paper page
  21. Schmid S. Clathrin-coated vesicle formation and protein sorting: an integrated process. Annu Rev Biochem. 1997;66:511–548. doi: 10.1146/annurev.biochem.66.1.511. [DOI] [PubMed] [Google Scholar] · DOI
  22. Sorkin A, Helin K, Waters CM, Carpenter G, Beguinot L. Multiple autophosphorylation sites of the epidermal growth factor receptor are essential for receptor kinase activity and internalization. J Biol Chem. 1992;267:8672–8678. [PubMed] [Google Scholar]
  23. Sorkin A, Mazzotti M, Sorkina T, Scotto L, Beguinot L. Epidermal growth factor interaction with clathrin adaptors is mediated by the Tyr974-containing internalization motif. J Biol Chem. 1996;271:13377–13384. doi: 10.1074/jbc.271.23.13377. [DOI] [PubMed] [Google Scholar] · DOI
  24. Stoorvogel W, Oorschot V, Geuze HJ. A novel class of clathrin-coated vesicles budding from endosomes. J Cell Biol. 1996;132:21–33. doi: 10.1083/jcb.132.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  25. Takei K, Mundigl O, Daniell L, De Camilli P. The synaptic vesicle cycle: a single vesicle budding step involving clathrin and dynamin. J Cell Biol. 1996;133:1237–1250. doi: 10.1083/jcb.133.6.1237. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  26. Traub LM, Kornfeld S. The trans-Golgi network: a late secretory sorting station. Curr Opin Cell Biol. 1997;9:527–533. doi: 10.1016/s0955-0674(97)80029-4. [DOI] [PubMed] [Google Scholar] · DOI
  27. Wang LH, Sudhof TC, Anderson RG. The appendage domain of alpha-adaptin is a high affinity binding site for dynamin. J Biol Chem. 1995;270:10079–10083. doi: 10.1074/jbc.270.17.10079. [DOI] [PubMed] [Google Scholar] · DOI
  28. Warren RA, Green FA, Stenberg PE, Enns CA. Distinct saturable pathways for the endocytosis of different tyrosine motifs. J Biol Chem. 1998;273:17056–17063. doi: 10.1074/jbc.273.27.17056. [DOI] [PubMed] [Google Scholar] · DOI
  29. Wendland B, Emr SD. Pan1p, yeast eps15, functions as a multivalent adaptor that coordinates protein-protein interactions essential for endocytosis. J Cell Biol. 1998;141:71–84. doi: 10.1083/jcb.141.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  30. Woodward MP, Roth TF. Coated vesicles: characterization, selective dissociation, and reassembly. Proc Natl Acad Sci USA. 1978;75:4394–4398. doi: 10.1073/pnas.75.9.4394. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  31. Yamabhai M, Hoffman NG, Hardison NL, McPherson PS, Castagnoli L, Cesareni G, Kay BK. Intersectin, a novel adaptor protein with two eps15 homology and five Src homology domains. J Biol Chem. 1998;273:31401–31407. doi: 10.1074/jbc.273.47.31401. [DOI] [PubMed] [Google Scholar] · DOI
  32. Ye W, Ali N, Bembenek ME, Shears SB, Lafer EM. Inhibition of clathrin assembly by high affinity binding of specific inositol polyphosphates to the synapse-specific clathrin assembly protein AP-3. J Biol Chem. 1995;270:1564–1568. [PubMed] [Google Scholar]
  33. Ye W, Lafer EM. Bacterially expressed F1–20/AP-3 assembles clathrin into cages with a narrow size distribution: implications for the regulation of quantal size during neurotransmission. J Neurosci Res. 1995a;41:15–26. doi: 10.1002/jnr.490410104. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  34. Ye W, Lafer EM. Clathrin binding and assembly activities of expressed domains of the synapse-specific clathrin assembly protein AP-3. J Biol Chem. 1995b;270:10933–10939. doi: 10.1074/jbc.270.18.10933. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  35. Zhang B, Koh YH, Beckstead RB, Budnik V, Ganetzky B, Bellen HJ. Synaptic vesicle size and number are regulated by a clathrin adaptor protein required for endocytosis. Neuron. 1998;21:1465–1475. doi: 10.1016/s0896-6273(00)80664-9. [DOI] [PubMed] [Google Scholar] · DOI · Paper page
  36. Zhou S, Sousa R, Tannery NH, Lafer EM. Characterization of novel synapse-specific protein. II. cDNA cloning and sequence analysis of the F1-20 protein. J Neurosci. 1992;12:2144–2155. doi: 10.1523/JNEUROSCI.12-06-02144.1992. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI

Authors
Tebar, F., Bohlander, S. K., & Sorkin, A.
Affiliations
  • University of Colorado Health (US)
  • University of Göttingen (DE)
ORCID
Venue
Molecular Biology of the Cell
Publisher
American Society for Cell Biology (ASCB)
Published
1999-08
Volume
10
Issue
8
Pages
2687-2702
ISSN
  • 1059-1524 (ISSN-L)
  • 1939-4586
DOI
10.1091/mbc.10.8.2687
PMC
PMC25500
OpenAlex
W2147149482
Cited by
320
References
37
Citations by year
2026: 7 · 2025: 4 · 2024: 6 · 2023: 12 · 2022: 9 · 2021: 10 · 2020: 10 · 2019: 15 · 2018: 8 · 2017: 8 · 2016: 14 · 2015: 9 · 2014: 26 · 2013: 16 · 2012: 25
FWCI
3.23
Type
journal article
Language
en
Open access
Locations
  • Molecular Biology of the Cell (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Dipòsit Digital de la Universitat de Barcelona (Universitat de Barcelona) (publishedVersion, cc-by-nc-sa)
  • RECERCAT (Consorci de Serveis Universitaris de Catalunya) (submittedVersion, cc-by-nc-sa)
Funders
  • Cancer League of Colorado
  • National Institutes of Health
  • National Cancer Institute (CA-46934, P30CA046934)
  • National Institute of Diabetes and Digestive and Kidney Diseases (R29DK046817, DK-46817)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Keywords
clathrin heavy chain, clathrin-coated pits, endocytosis, mannose 6-phosphate receptor, trans-Golgi network, plasma membrane
MeSH terms
Animals, Biological Transport, Clathrin, Coated Pits, Cell-Membrane, Endocytosis, Golgi Apparatus, Humans, Luminescent Proteins, Nerve Tissue Proteins, Phosphoproteins, Endosomes, Recombinant Proteins, COS Cells, Adaptor Proteins, Vesicular Transport, Monomeric Clathrin Assembly Proteins, Green Fluorescent Proteins, ErbB Receptors

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