Home LiteratureArticle Details
PMID: 19581412 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Quantitative proteomics identifies a Dab2/integrin module regulating cell migration.

The Journal of cell biology ·Vol. 186 ·No. 1 ·2009-07-13 ·Pages 99-111

Teckchandani A, Toida N, Goodchild J, Henderson C, Watts J, Wollscheid B, Cooper JA

Abstract

Clathrin-associated endocytic adapters recruit cargoes to coated pits as a first step in endocytosis. We developed an unbiased quantitative proteomics approach to identify and quantify glycoprotein cargoes for an endocytic adapter, Dab2. Surface levels of integrins beta1, alpha1, alpha2, and alpha3 but not alpha5 or alphav chains were specifically increased on Dab2-deficient HeLa cells. Dab2 colocalizes with integrin beta1 in coated pits that are dispersed over the cell surface, suggesting that it regulates bulk endocytosis of inactive integrins. Depletion of Dab2 inhibits cell migration and polarized movement of integrin beta1 and vinculin to the leading edge. By manipulating intracellular and surface integrin beta1 levels, we show that migration speed correlates with the intracellular integrin pool but not the surface level. Together, these results suggest that Dab2 internalizes integrins freely diffusing on the cell surface and that Dab2 regulates migration, perhaps by maintaining an internal pool of integrins that can be recycled to create new adhesions at the leading edge.

MeSH Terms
Adaptor Protein Complex 2/metabolism Adaptor Proteins, Signal Transducing/deficiency,metabolism Animals Apoptosis Regulatory Proteins Cell Membrane/metabolism Cell Movement Clathrin/metabolism Endocytosis HeLa Cells Humans Integrin alpha1/metabolism Integrin beta1/metabolism Integrins/metabolism Membrane Proteins/metabolism Mice Nerve Tissue Proteins/metabolism Protein Transport Proteomics Tumor Suppressor Proteins
Chemicals
Adaptor Protein Complex 2 Adaptor Proteins, Signal Transducing Apoptosis Regulatory Proteins Clathrin DAB2 protein, human Integrin alpha1 Integrin beta1 Integrins Membrane Proteins Nerve Tissue Proteins Numb protein, human Tumor Suppressor Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Teckchandani Anjali
Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Toida Natalie
Goodchild Jake
Henderson Christine
Watts Julian
Wollscheid Bernd
Cooper Jonathan A
References (63)
63 references, click to expand
  1. Multisite phosphorylation by Cdk2 and GSK3 controls cyclin E degradation.
    Mol Cell. 2003 Aug;12(2):381-92 PMID: 14536078
  2. Mouse numb is an essential gene involved in cortical neurogenesis.
    Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6844-9 PMID: 10841580
  3. Integrin trafficking and the control of cell migration.
    Traffic. 2006 Jan;7(1):14-21 PMID: 16445683
  4. Antibody against human alpha 1 beta 1 integrin inhibits HeLa cell adhesion to laminin and to type I, IV, and V collagens.
    Biochem Biophys Res Commun. 1995 Apr 6;209(1):205-12 PMID: 7726837
  5. Integrin trafficking regulated by Rab21 is necessary for cytokinesis.
    Dev Cell. 2008 Sep;15(3):371-385 PMID: 18804435
  6. Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics.
    Mol Cell Proteomics. 2002 May;1(5):376-86 PMID: 12118079
  7. Phosphorylation of ACAP1 by Akt regulates the stimulation-dependent recycling of integrin beta1 to control cell migration.
    Dev Cell. 2005 Nov;9(5):663-73 PMID: 16256741
  8. Regulation of human B-cell precursor adhesion to bone marrow stromal cells by cytokines that exert opposing effects on the expression of vascular cell adhesion molecule-1 (VCAM-1).
    Blood. 1993 May 1;81(9):2272-82 PMID: 7683214
  9. Stimulation-dependent recycling of integrin beta1 regulated by ARF6 and Rab11.
    Traffic. 2004 Jan;5(1):20-36 PMID: 14675422
  10. Microtubule targeting of substrate contacts promotes their relaxation and dissociation.
    J Cell Biol. 1999 Sep 6;146(5):1033-44 PMID: 10477757
  11. Focal adhesion disassembly requires clathrin-dependent endocytosis of integrins.
    FEBS Lett. 2009 Apr 17;583(8):1337-43 PMID: 19306879
  12. DOC-2/hDab-2 inhibits ILK activity and induces anoikis in breast cancer cells through an Akt-independent pathway.
    Oncogene. 2001 Oct 18;20(47):6960-4 PMID: 11687976
  13. Protein kinase B/Akt acts via glycogen synthase kinase 3 to regulate recycling of alpha v beta 3 and alpha 5 beta 1 integrins.
    Mol Cell Biol. 2004 Feb;24(4):1505-15 PMID: 14749368
  14. Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase.
    Nat Cell Biol. 2005 Jun;7(6):581-90 PMID: 15895076
  15. Characterization of a novel negative regulator (DOC-2/DAB2) of c-Src in normal prostatic epithelium and cancer.
    J Biol Chem. 2003 Feb 28;278(9):6936-41 PMID: 12473651
  16. Restoration of positioning control following Disabled-2 expression in ovarian and breast tumor cells.
    Oncogene. 2000 Oct 5;19(42):4847-54 PMID: 11039902
  17. Circulating integrins: alpha 5 beta 1, alpha 6 beta 4 and Mac-1, but not alpha 3 beta 1, alpha 4 beta 1 or LFA-1.
    EMBO J. 1992 Feb;11(2):405-10 PMID: 1531629
  18. Dab2 regulates clathrin assembly and cell spreading.
    Biochem J. 2009 Mar 15;418(3):701-15 PMID: 19000037
  19. Disabled-2 (Dab2) mediates transforming growth factor beta (TGFbeta)-stimulated fibronectin synthesis through TGFbeta-activated kinase 1 and activation of the JNK pathway.
    J Biol Chem. 2005 Jul 8;280(27):25920-7 PMID: 15894542
  20. Clustering induces a lateral redistribution of alpha 2 beta 1 integrin from membrane rafts to caveolae and subsequent protein kinase C-dependent internalization.
    Mol Biol Cell. 2004 Feb;15(2):625-36 PMID: 14657242
  21. The alphavbeta5 integrin functions as an endocytic receptor for vitronectin.
    J Cell Sci. 1998 Feb;111 ( Pt 4):425-33 PMID: 9443892
  22. Disabled-2 exhibits the properties of a cargo-selective endocytic clathrin adaptor.
    EMBO J. 2002 Sep 16;21(18):4915-26 PMID: 12234931
  23. Internalization of the alpha 5 beta 1 integrin does not depend on "NPXY" signals.
    Biochem Biophys Res Commun. 1994 Mar 15;199(2):603-11 PMID: 7510958
  24. Clathrin-coated vesicle formation and protein sorting: an integrated process.
    Annu Rev Biochem. 1997;66:511-48 PMID: 9242916
  25. An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database.
    J Am Soc Mass Spectrom. 1994 Nov;5(11):976-89 PMID: 24226387
  26. A single common portal for clathrin-mediated endocytosis of distinct cargo governed by cargo-selective adaptors.
    Mol Biol Cell. 2006 Oct;17(10):4300-17 PMID: 16870701
  27. Ca(2+)- and calcineurin-dependent recycling of an integrin to the front of migrating neutrophils.
    Nature. 1995 Sep 7;377(6544):75-9 PMID: 7544874
  28. Cargo recognition during clathrin-mediated endocytosis: a team effort.
    Curr Opin Cell Biol. 2004 Aug;16(4):392-9 PMID: 15261671
  29. Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells.
    J Cell Biol. 2001 Jun 25;153(7):1427-40 PMID: 11425873
  30. Cargo and dynamin regulate clathrin-coated pit maturation.
    PLoS Biol. 2009 Mar 17;7(3):e57 PMID: 19296720
  31. Endocytic recycling pathways: emerging regulators of cell migration.
    Curr Opin Cell Biol. 2006 Oct;18(5):549-57 PMID: 16904305
  32. Restriction of Src activity by Cullin-5.
    Curr Biol. 2009 Jan 27;19(2):157-62 PMID: 19147357
  33. The adaptor protein Dab2 sorts LDL receptors into coated pits independently of AP-2 and ARH.
    J Cell Sci. 2006 Oct 15;119(Pt 20):4235-46 PMID: 16984970
  34. Sorting it out: AP-2 and alternate clathrin adaptors in endocytic cargo selection.
    J Cell Biol. 2003 Oct 27;163(2):203-8 PMID: 14581447
  35. A tyrosine-based sorting signal in the beta2 integrin cytoplasmic domain mediates its recycling to the plasma membrane and is required for ligand-supported migration.
    EMBO J. 1999 Sep 15;18(18):4915-25 PMID: 10487744
  36. PDGF-regulated rab4-dependent recycling of alphavbeta3 integrin from early endosomes is necessary for cell adhesion and spreading.
    Curr Biol. 2001 Sep 18;11(18):1392-402 PMID: 11566097
  37. Numb controls integrin endocytosis for directional cell migration with aPKC and PAR-3.
    Dev Cell. 2007 Jul;13(1):15-28 PMID: 17609107
  38. Integrin avidity regulation: are changes in affinity and conformation underemphasized?
    Curr Opin Cell Biol. 2003 Oct;15(5):547-56 PMID: 14519389
  39. Mass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins.
    Nat Biotechnol. 2009 Apr;27(4):378-86 PMID: 19349973
  40. Endocytosis and recycling of the fibronectin receptor in CHO cells.
    EMBO J. 1989 May;8(5):1341-8 PMID: 2527741
  41. FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly.
    Nat Cell Biol. 2004 Feb;6(2):154-61 PMID: 14743221
  42. Morphogenesis of human endothelial cells is inhibited by DAB2 via Src.
    FEBS Lett. 2008 Jul 23;582(17):2542-8 PMID: 18582465
  43. Moving membrane up to the front of migrating cells.
    Cell. 1996 May 17;85(4):465-7 PMID: 8653781
  44. Disabled-2 (Dab2) is required for transforming growth factor beta-induced epithelial to mesenchymal transition (EMT).
    J Biol Chem. 2005 Apr 29;280(17):17540-8 PMID: 15734730
  45. Fibronectin receptor exhibits high lateral mobility in embryonic locomoting cells but is immobile in focal contacts and fibrillar streaks in stationary cells.
    J Cell Biol. 1988 Oct;107(4):1385-96 PMID: 2971668
  46. Depletion of intracellular potassium disrupts coated pits and reversibly inhibits cell polarization during fibroblast spreading.
    J Cell Biol. 1993 Mar;120(6):1449-59 PMID: 8449988
  47. Endocytosis of megalin by visceral endoderm cells requires the Dab2 adaptor protein.
    J Cell Sci. 2005 Nov 15;118(Pt 22):5345-55 PMID: 16263760
  48. Oriented endocytic recycling of alpha5beta1 in motile neutrophils.
    Blood. 2000 Apr 15;95(8):2471-80 PMID: 10753823
  49. Cycling of cell-surface MHC glycoproteins through primaquine-sensitive intracellular compartments.
    Nature. 1990 Aug 16;346(6285):655-7 PMID: 2166918
  50. Integrin-ligand binding properties govern cell migration speed through cell-substratum adhesiveness.
    Nature. 1997 Feb 6;385(6616):537-40 PMID: 9020360
  51. Disabled-2 colocalizes with the LDLR in clathrin-coated pits and interacts with AP-2.
    Traffic. 2001 Feb;2(2):111-23 PMID: 11247302
  52. Talin binding to integrin beta tails: a final common step in integrin activation.
    Science. 2003 Oct 3;302(5642):103-6 PMID: 14526080
  53. A statistical model for identifying proteins by tandem mass spectrometry.
    Anal Chem. 2003 Sep 1;75(17):4646-58 PMID: 14632076
  54. p96, a MAPK-related protein, is consistently downregulated during mouse mammary carcinogenesis.
    Oncogene. 1998 Sep 3;17(9):1173-8 PMID: 9764828
  55. Integrin beta cytoplasmic domain interactions with phosphotyrosine-binding domains: a structural prototype for diversity in integrin signaling.
    Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2272-7 PMID: 12606711
  56. Integrins: versatility, modulation, and signaling in cell adhesion.
    Cell. 1992 Apr 3;69(1):11-25 PMID: 1555235
  57. Disabled-2 (Dab2) is an SH3 domain-binding partner of Grb2.
    Oncogene. 1998 Mar 26;16(12):1561-9 PMID: 9569023
  58. Dual roles for the Dab2 adaptor protein in embryonic development and kidney transport.
    EMBO J. 2002 Apr 2;21(7):1555-64 PMID: 11927540
  59. Real-time analysis of clathrin-mediated endocytosis during cell migration.
    J Cell Sci. 2003 Mar 1;116(Pt 5):847-55 PMID: 12571282
  60. Genetic analysis of beta1 integrin "activation motifs" in mice.
    J Cell Biol. 2006 Sep 11;174(6):889-99 PMID: 16954348
  61. Automated statistical analysis of protein abundance ratios from data generated by stable-isotope dilution and tandem mass spectrometry.
    Anal Chem. 2003 Dec 1;75(23):6648-57 PMID: 14640741
  62. Molecular cloning of differentially expressed genes in human epithelial ovarian cancer.
    Gynecol Oncol. 1994 Feb;52(2):247-52 PMID: 8314147
  63. Cargo regulates clathrin-coated pit dynamics.
    Cell. 2006 Oct 6;127(1):113-24 PMID: 17018281
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.200812160
Published
2009-07-13
Epub
2009-00-06
Pages
99-111
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2712992
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066257 · United States
NIAID NIH HHS · R01 AI051344 · United States
NIGMS NIH HHS · R01-GM66257 · United States
NHLBI NIH HHS · N01HV28179 · United States
NHLBI NIH HHS · N01-HV-28179-22 · United States
NIGMS NIH HHS · GM078776 · United States
NIGMS NIH HHS · F32 GM078776 · United States
NIAID NIH HHS · R01-AI51344-01 · United States
External Links
PubMed source DOI: 10.1083/jcb.200812160 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: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2