CoChat
Sign Up
Cited by 57Cellular transport and secretionCaveolin-1 and cellular processesBiotin and Related Studies

AAGAB Controls AP2 Adaptor Assembly in Clathrin-Mediated Endocytosis

Open access
Gulbranson, Daniel R., Crisman, Lauren, Lee, MyeongSeon, Ouyang, Yan, Menasche, Bridget L., Demmitt, Brittany A., Wan, Chun, Nomura, Toshifumi +3 more·2019·Developmental Cell
DOI 10.1016/j.devcel.2019.06.013PMC PMC6702059
Open this paper in CoChat — free accountOpen PDF

Abstract

Multimeric adaptors are broadly involved in vesicle-mediated membrane trafficking. AP2 adaptor, in particular, plays a central role in clathrin-mediated endocytosis (CME) by recruiting cargo and clathrin to endocytic sites. It is generally thought that trafficking adaptors such as AP2 adaptor assemble spontaneously. In this work, however, we discovered that AP2 adaptor assembly is an ordered process controlled by alpha and gamma adaptin binding protein (AAGAB), an uncharacterized factor identified in our genome-wide genetic screen of CME. AAGAB guides the sequential association of AP2 subunits and stabilizes assembly intermediates. Without the assistance of AAGAB, AP2 subunits fail to form the adaptor complex, leading to their degradation. The function of AAGAB is abrogated by a mutation that causes punctate palmoplantar keratoderma type 1 (PPKP1), a human skin disease. Since other multimeric trafficking adaptors operate in an analogous manner to AP2 adaptor, their assembly likely involves a similar regulatory mechanism.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.devcel.2019.06.013

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Top-ranking binding proteins of Reps1 included subunits of the AP2 adaptor complex.

    “Top-ranking binding proteins of Reps1 included subunits of the AP2 adaptor complex (Fig. 3B and table S4), a key regulator of CME required for GLUT-SPR internalization (21).”

    From Regulation of cargo exocytosis by a Reps1-Ralbp1-RalA module (2023) · cites this paper as [21]

    The citation is attached to the description of AP2, not to the citing paper’s own finding that Reps1 binds AP2 subunits. The Introduction identifies AP2 as the predominant clathrin adaptor in CME, and the Results describe CME-mediated internalization of an HA-GLUT4-GFP reporter. Our copy does not establish that AP2 is required for internalization of the specifically named GLUT-SPR reporter. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • GLUT4 depends on general exocytic and endocytic pathways to regulate its surface levels.

    “The reporter is based on the glucose transporter GLUT4 (Fig. 1A), which depends on general exocytic and endocytic pathways to regulate its surface levels (3, 19–21).”

    From Regulation of cargo exocytosis by a Reps1-Ralbp1-RalA module (2023) · cites this paper as [21]

    The Results section (‘Identification of CME regulators in a genome-wide genetic screen’; page not provided) says that the GLUT4 reporter, like native GLUT4, is continuously internalized by clathrin-mediated endocytosis, and describes measuring its surface levels. The passages found do not establish the claim’s exocytic-pathway component; GLUT4 exocytosis appears in a cited work’s title in the References, not as a finding stated here. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 46

  1. Antonescu CN, McGraw TE, and Klip A (2014). Reciprocal regulation of endocytosis and metabolism. Cold Spring Harbor perspectives in biology 6, a016964. 24984778 PMC4067987
  2. Bogan JS, and Kandror KV (2010). Biogenesis and regulation of insulin-responsive vesicles containing GLUT4. Current opinion in cell biology 22, 506– 512. 20417083 PMC2910140
  3. Boll W, Ohno H, Songyang Z, Rapoport I, Cantley LC, Bonifacino JS, and Kirchhausen T (1996). Sequence requirements for the recognition of tyrosine-based endocytic signals by clathrin AP-2 complexes. EMBO J 15, 5789– 5795. 8918456 PMC452326
  4. Bonifacino JS, and Glick BS (2004). The mechanisms of vesicle budding and fusion. Cell 116, 153– 166. 14744428
  5. Brodsky FM (2012). Diversity of clathrin function: new tricks for an old protein. Annu Rev Cell Dev Biol 28, 309– 336. 22831640
  6. Collins BM, McCoy AJ, Kent HM, Evans PR, and Owen DJ (2002). Molecular architecture and functional model of the endocytic AP2 complex. Cell 109, 523– 535. 12086608
  7. Conner SD, and Schmid SL (2003). Differential requirements for AP-2 in clathrin-mediated endocytosis. The Journal of cell biology 162, 773– 779. 12952931 PMC2172816
  8. Di Paolo G, and De Camilli P (2006). Phosphoinositides in cell regulation and membrane dynamics. Nature 443, 651– 657. 17035995
  9. Doray B, Lee I, Knisely J, Bu G, and Kornfeld S (2007). The gamma/sigma1 and alpha/sigma2 hemicomplexes of clathrin adaptors AP-1 and AP-2 harbor the dileucine recognition site. Molecular biology of the cell 18, 1887– 1896. 17360967 PMC1855031
  10. Fotin A, Cheng Y, Sliz P, Grigorieff N, Harrison SC, Kirchhausen T, and Walz T (2004). Molecular model for a complete clathrin lattice from electron cryomicroscopy. Nature 432, 573– 579. 15502812
  11. Giehl KA, Eckstein GN, Pasternack SM, Praetzel-Wunder S, Ruzicka T, Lichtner P, Seidl K, Rogers M, Graf E, Langbein L, (2012). Nonsense Mutations in AAGAB Cause Punctate Palmoplantar Keratoderma Type Buschke-Fischer-Brauer. American Journal of Human Genetics 91, 754– 759. 23000146 PMC3484507
  12. Gorynia S, Lorenz TC, Costaguta G, Daboussi L, Cascio D, and Payne GS (2012). Yeast Irc6p is a novel type of conserved clathrin coat accessory factor related to small G proteins. Molecular biology of the cell 23, 4416– 4429. 22993212 PMC3496615
  13. Gulbranson DR, Davis EM, Demmitt BA, Ouyang Y, Ye Y, Yu H, and Shen J (2017). RABIF/MSS4 is a Rab-stabilizing holdase chaperone required for GLUT4 exocytosis. Proc Natl Acad Sci U S A 114(39), E8224– E8233. 28894007 PMC5625932
  14. Jackson LP, Kelly BT, McCoy AJ, Gaffry T, James LC, Collins BM, Honing S, Evans PR, and Owen DJ (2010). A large-scale conformational change couples membrane recruitment to cargo binding in the AP2 clathrin adaptor complex. Cell 141, 1220– 1229. 20603002 PMC3655264
  15. Jewell JL, Oh E, and Thurmond DC (2010). Exocytosis mechanisms underlying insulin release and glucose uptake: conserved roles for Munc18c and syntaxin 4. Am J Physiol Regul Integr Comp Physiol 298, R517– 531. 20053958 PMC2838661
  16. Kaksonen M, and Roux A (2018). Mechanisms of clathrin-mediated endocytosis. Nature reviews Molecular cell biology 19, 313– 326. 29410531
  17. Kaksonen M, Toret CP, and Drubin DG (2006). Harnessing actin dynamics for clathrin-mediated endocytosis. Nature reviews Molecular cell biology 7, 404– 414. 16723976
  18. Kelly BT, Graham SC, Liska N, Dannhauser PN, Honing S, Ungewickell EJ, and Owen DJ (2014). Clathrin adaptors. AP2 controls clathrin polymerization with a membrane-activated switch. Science (New York, NY) 345, 459– 463. PMC4333214 25061211
  19. Kelly BT, McCoy AJ, Spate K, Miller SE, Evans PR, Honing S, and Owen DJ (2008). A structural explanation for the binding of endocytic dileucine motifs by the AP2 complex. Nature 456, 976– 979. 19140243 PMC4340503
  20. Kim YE, Hipp MS, Bracher A, Hayer-Hartl M, and Hartl FU (2013). Molecular chaperone functions in protein folding and proteostasis. Annual review of biochemistry 82, 323– 355. 23746257
  21. Kirchhausen T, Owen D, and Harrison SC (2014). Molecular structure, function, and dynamics of clathrin-mediated membrane traffic. Cold Spring Harbor perspectives in biology 6, a016725. 24789820 PMC3996469
  22. Li W, Xu H, Xiao T, Cong L, Love MI, Zhang F, Irizarry RA, Liu JS, Brown M, and Liu XS (2014). MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens. Genome biology 15, 554. 25476604 PMC4290824
  23. McMahon HT, and Boucrot E (2011). Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nature reviews Molecular cell biology 12, 517– 533. 21779028
  24. Menasche BL, Crisman L, Gulbranson DR, Davis EM, Yu H, and Shen J (2018). Fluorescence Activated Cell Sorting (FACS) in Genome-Wide Genetic Screening of Membrane Trafficking. Curr Protoc Cell Biol, e68. 30265447 PMC6386612
  25. Mettlen M, Chen PH, Srinivasan S, Danuser G, and Schmid SL (2018). Regulation of Clathrin-Mediated Endocytosis. Annu Rev Biochem. PMC6383209 29661000
  26. Motley A, Bright NA, Seaman MN, and Robinson MS (2003). Clathrin-mediated endocytosis in AP-2-depleted cells. The Journal of cell biology 162, 909– 918. 12952941 PMC2172830
  27. Muretta JM, Romenskaia I, and Mastick CC (2008). Insulin releases Glut4 from static storage compartments into cycling endosomes and increases the rate constant for Glut4 exocytosis. J Biol Chem 283, 311– 323. 17967900
  28. Nomura T, Yoneta A, Pohler E, Suzuki S, Osawa R, Mizuno O, Ohguchi Y, Nomura Y, Yamashita T, McLean WHI, (2015). Punctate Palmoplantar Keratoderma Type 1: A Novel AAGAB Mutation and Efficacy of Etretinate. Acta Derm-Venereol 95, 110– 111. 24573067
  29. Ohno H, Stewart J, Fournier MC, Bosshart H, Rhee I, Miyatake S, Saito T, Gallusser A, Kirchhausen T, and Bonifacino JS (1995). Interaction of tyrosine-based sorting signals with clathrin-associated proteins. Science (New York, NY) 269, 1872– 1875. 7569928
  30. Page LJ, Sowerby PJ, Lui WW, and Robinson MS (1999). Gamma-synergin: an EH domain-containing protein that interacts with gamma-adaptin. The Journal of cell biology 146, 993– 1004. 10477754 PMC2169493
  31. Pearse BM, and Robinson MS (1984). Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin. EMBO J 3, 1951– 1957. 6149117 PMC557627
  32. Pohler E, Mamai O, Hirst J, Zamiri M, Horn H, Nomura T, Irvine AD, Moran B, Wilson NJ, Smith FJ, (2012). Haploinsufficiency for AAGAB causes clinically heterogeneous forms of punctate palmoplantar keratoderma. Nat Genet 44, 1272– 1276. 23064416 PMC3836166
  33. Ramanan V, Agrawal NJ, Liu J, Engles S, Toy R, and Radhakrishnan R (2011). Systems biology and physical biology of clathrin-mediated endocytosis. Integr Biol (Camb) 3, 803– 815. 21792431 PMC3153420
  34. Rathore SS, Bend EG, Yu H, Hammarlund M, Jorgensen EM, and Shen J (2010). Syntaxin N-terminal peptide motif is an initiation factor for the assembly of the SNARE-Sec1/Munc18 membrane fusion complex. Proc Natl Acad Sci U S A 107, 22399– 22406. 21139055 PMC3012463
  35. Ren X, Farias GG, Canagarajah BJ, Bonifacino JS, and Hurley JH (2013). Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell 152, 755– 767. 23415225 PMC3913725
  36. Rothman JE (1994). Mechanisms of intracellular protein transport. Nature 372, 55– 63. 7969419
  37. Rowland AF, Fazakerley DJ, and James DE (2011). Mapping Insulin/GLUT4 Circuitry. Traffic 12, 672– 681. 21401839
  38. Sanjana NE, Shalem O, and Zhang F (2014). Improved vectors and genome-wide libraries for CRISPR screening. Nature methods 11, 783– 784. 25075903 PMC4486245
  39. Schekman R, and Novick P (2004). 23 genes, 23 years later. Cell 116, S13– 15, 11 p following S19. 15055575
  40. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, (2014). Genome-scale CRISPR-Cas9 knockout screening in human cells. Science (New York, NY) 343, 84– 87. PMC4089965 24336571
  41. Shen J, Chen X, Hendershot L, and Prywes R (2002). ER stress regulation of ATF6 localization by dissociation of BiP/GRP78 binding and unmasking of Golgi localization signals. Developmental cell 3, 99– 111. 12110171
  42. Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, Sabatini DM, Chen IS, Hahn WC, Sharp PA, (2003). Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9, 493– 501. 12649500 PMC1370415
  43. Traub LM, and Bonifacino JS (2013). Cargo recognition in clathrin-mediated endocytosis. Cold Spring Harbor perspectives in biology 5, a016790. 24186068 PMC3809577
  44. Yu A, Xing Y, Harrison SC, and Kirchhausen T (2010). Structural analysis of the interaction between Dishevelled2 and clathrin AP-2 adaptor, a critical step in noncanonical Wnt signaling. Structure 18, 1311– 1320. 20947020 PMC2992793
  45. Yu H, Crisman L, Stowell MHBand Shen J (2019). Functional Reconstitution of Intracellular Vesicle Fusion Using Purified SNAREs and Sec1/Munc18 (SM) Proteins. Methods Mol Biol I860, 237– 249. PMC6869331 30317509
  46. Yu H, Rathore SS, Shen C, Liu Y, Ouyang Y, Stowell MH, and Shen J (2015). Reconstituting Intracellular Vesicle Fusion Reactions: The Essential Role of Macromolecular Crowding. J Am Chem Soc 137, 12873– 12883. 26431309 PMC4638180

Cited by

Sorted by citations

Authors
Gulbranson, D. R., Crisman, L., Lee, M., Ouyang, Y., Menasche, B. L., Demmitt, B. A., Wan, C., Nomura, T., Ye, Y., Yu, H., & Shen, J.
Affiliations
  • University of Colorado Boulder (US)
  • Hokkaido University (JP)
  • National Institutes of Health (US)
  • Nanjing Normal University (CN)
Corresponding author
  • Yu, Haijia
  • Shen, Jingshi
ORCID
Venue
Developmental Cell
Publisher
Elsevier BV
Published
2019-08
Volume
50
Issue
4
Pages
436-446.e5
ISSN
1534-5807 (ISSN-L)
DOI
10.1016/j.devcel.2019.06.013
PMC
PMC6702059
OpenAlex
W2962882614
Cited by
57
References
1
Citations by year
2026: 9 · 2025: 9 · 2024: 6 · 2023: 8 · 2022: 9 · 2021: 5 · 2020: 9 · 2019: 2
FWCI
3.28
Type
journal article
Language
en
Open access
Licence
Locations
  • Developmental Cell (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Aerospace Science Foundation of China
  • National Institutes of Health (DK095367, GM126960)
  • University of Colorado
  • American Heart Association (GM088759)
  • National Natural Science Foundation of China (31871425, 91854117)
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
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
Keywords
clathrin-mediated endocytosis, protein degradation, genome-wide screen
MeSH terms
Amino Acid Sequence, Cell Membrane, Clathrin, Endocytosis, Humans, Keratoderma, Palmoplantar, Protein Binding, Protein Transport, Adaptor Proteins, Vesicular Transport, Adaptor Protein Complex 2, Proteolysis

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review