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Interchangeable alpha chain cytoplasmic domains play a positive role in control of cell adhesion mediated by VLA-4, a beta 1 integrin



Integrins can exist in a range of functional states, depending on the cell type and its state of activation. Although the mechanism that controls activity is unknown, it has been suggested that for some integrins, alpha chain cytoplasmic domains may exert either a negative effect or no effect on adhesion function. To address this issue for VLA- 4 (an alpha 4 beta 1 heterodimer), we constructed an alpha 4 cytoplasmic deletion mutant and chimeric alpha chains composed of the extracellular domains of alpha 4 and the cytoplasmic domains of alpha 2, alpha 4, or alpha 5. Upon stable transfection of wild-type alpha 4, VLA-4 heterodimer was obtained that mediated (a) poor adhesion to CS1 peptide, fibronectin, or vascular cell adhesion molecule 1 (VCAM-1) (in K562 cells); (b) poor adhesion to CS1 peptide but moderate adhesion to VCAM-1 (in MIP101 cells); and (c) moderate adhesion to both CS1 peptide and VCAM-1 (in PMWK cells). Chimeric alpha 4 constructs and wild-type alpha 4 yielded similar results in these cell lines. In contrast, truncation of the alpha 4 cytoplasmic domain (after the conserved GFFKR motif) caused an almost complete loss of adhesive activity in all three cell lines. Thus, several interchangeable alpha chain cytoplasmic domains play a fundamentally positive role in determining the state of constitutive activity for VLA-4. The alpha chain cytoplasmic domain is also required for agonist-stimulated adhesion, since phorbol ester stimulated the cell adhesion mediated by wild-type and chimeric alpha chains, but not by the cytoplasmic deletion mutant. The inactivity of both wild-type VLA-4 (in K562 cells), and truncated VLA-4 (in all three cell lines) was overcome by the addition of a stimulatory anti-beta 1 monoclonal antibody. Thus, the alpha cytoplasmic domain-dependent cellular mechanism controlling both constitutive and agonist-stimulated VLA-4 activity could be bypassed by external manipulation of the integrin

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  1. (1991). 31 integrin-mediated lymphocyte adherence to extracellular matrix is enhanced by phorbol ester treatment.
  2. (1993). A high af~nity conformational state on FLA integrin heterodimers induced by an anti-~l chain monoclonal antibody.
  3. (1992). A monoclonal antibody to 131 integrin (CD29) stimulates VLA-dependent adherence of leukocytes to human umbilical vein endothelial cells and matrix components.
  4. (1989). A simple method for site directed mutagenesis using the polymerase chain reaction.
  5. (1989). Adhesion of human T-lymphoid cells to fibronectin is mediated by two different fibronectin domains.
  6. (1991). Adhesion of immature thymocytes to thymic stromal cells through fibronectin molecules and its significance for the induction of thymocyte differentiation.
  7. (1992). Adhesion to VCAM-1 and fibronectin: comparison of c~4B1 (VLA-4) and ol4/37 on the human cell line JY.
  8. (1991). Adhesive properties of the B3 integrins: comparison of GP IIb-IIIa and the vitronectin receptor individually expressed in human melanoma cells.J.
  9. (1990). Affinity modulation of the czlrd33 integrin (platelet GPlIb-IIIa) is an intrinsic property of the receptor. Cell Regul.
  10. Afiinity modulation of integrin a5131: regulation of the functional response by soluble fibronectin.J.
  11. (1989). Analysis of the role of glycosylation of the human fibronectin receptor.
  12. (1988). Attachment, spreading and locomotion of avian neural crest cells are mediated by multiple adhesion sites on fibronectin molecules.
  13. (1991). Calcium as a potent physiological regulator ofintegrin-mediated cell adhesion.
  14. (1991). Cell migration and actin organization in cultured human primary, recurrent cutaneous and metastatic mdanoma.
  15. (1989). Cellular adhesion: GPIIb/IIIa as a prototypic adhesion receptor.
  16. (1983). Characterization of a novel differentiation antigen complex recognized by a monoclonal antibody (A-1A5): unique activation-specific molecular forms on stimulated T cells, f Immunol.
  17. (1987). Characterization of the cell surface heterodimer VLA-4 and related peptides.
  18. (1990). Constitutive and induced phosphorylation of the c~- and B-chains of the CDll/CD18 leukocyte integrin family.J.
  19. (1989). Constitutive and stimulus-induced phosphorylation of CD11/CD18 leukocyte adhesion molecules.
  20. (1992). Distinct cellular functions mediated by different VLA integrin cr subunit cytoplasmic domains.
  21. (1988). Dynamic membranecytoskeletal interactions: specific association of integrin and talin arises in vivo after phorbol ester treatment of peripheral blood lymphocytes.
  22. (1991). Effect of a new monoclonal antibody, TA-2, that inhibits lymphocyte adherence to cytokine stimulated endothelium in the rat.J.
  23. (1989). Enhancement of LFA-1-mediated cell adhesion by triggering through CD2 or CD3 on T lymphocytes.
  24. (1991). Evidence for a role of the integrin VLA-4 in lymphohemopoisis.
  25. (1990). Expression and function of chicken integrin beta-1 subunit and its cytoplasmic domain mutants
  26. (1991). Expression and functional char659 Kassner and Hemler acterization of a soluble form of vascular cell adhesion molecule 1 (VCAM1).
  27. (1992). Expression and ligand-binding of the integrin alpha 4 beta a (VLA-4) on neuralcrest-derived tumor cell lines.
  28. (1991). Fibronectin and VLA-4 in haematopoietic stem cell-microenvironment interactions.
  29. (1992). Functional and structural analysis of VLA-4 integrin ~4 subunit cleavage.
  30. Glycoproteins of 210,000 and 130,000 m.w. on activated T cells: cell distribution and antigenic relation to components on resting cells and T cell lines.
  31. (1991). GPIIbIIIa: the responsive integrin.
  32. (1989). Identification and characterization of the lymphocyte adhesion receptor for an alternative cell attachment domain in plasma fibronectin.J.
  33. (1990). Identification of surface proteins mediating adherence of CDll/CD18-deficient lymphoblastoid cells to cultured human endothelium, j.
  34. (1992). Increased binding of synovial T lymphocytes from rheumatoid arthritis to endothelialleukocyte adhesion molecule-1 (ELAM-1) and vascular cell adhesion molecule-1 (VCAM-1).
  35. (1990). Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes: a CD11/CD18-independent adhesion mechanism.
  36. (1987). Induction of the fibrinogen receptor on human platelets by intracellular mediators.J.
  37. (1992). Integrin modulating factor-l: a lipid that alters the function of leukocyte integrins.
  38. (1992). Integrins: versatility, modulation and signalling in cell adhesion.
  39. is0: a novel form of the integrin c~ 4 subunit.J.
  40. (1990). Lymphoid cells recognize an alternatively spliced segment of fibronectin via the integrin receptor c~4B1.
  41. (1984). Lynphocyte activation antigens I. A monoclonal antibody, antiAct I, defines a new late lymphocyte activation antigen.J.
  42. (1988). Macrophage interactions with laminin: PMA selectively induces the adherence and spreading of mouse macrophages on a laminin substratum.
  43. (1991). Mechanisms of eosinophil adherence to cultured vascular endothelial cells. Eosinophils bind to the cytokine-induced endothelial ligand vascular cell adhesion molecule-1 via the very late activation antigen-4 integrin receptor.
  44. (1991). Modulation of the affinity of integrin cfiIBB3 (GPIIb-IIIa) by the cytoplasmic domain of cflIb. Science (Wash.
  45. (1990). Modulation ofvitronectin receptor binding by membrane lipid composition.J.
  46. (1982). Monoclonal antibodies reacting with immunogenic mycophsma proteins present in human hematopoietic cell lines.
  47. (1993). Multiple activation states of VLA-4: mechanistic differences between adhesion to CS1/fibronectin and to VCAM-1.
  48. (1993). Multiple functional forms of the integrin VLA-2 derived from a single od cDNA clone: interconversion of forms induced by an anti-ill antibody.
  49. (1989). Phorbol ester modulation of integrin-mediated cell adhesion: a postreceptor event.
  50. (1986). Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes.
  51. (1992). Prevention of experimental autoimmune encephalomyelitis by antibodies against a4B1 integrin.
  52. (1987). Primary structure of human plasma fibronectin: characterization of a 38 kDa domain containing C-terminal heparin-binding site (Hep III site) and a region of molecular heterogeneity.
  53. (1985). Reconstitution of an active surface T3/T-cell antigen receptor by DNA transfer.
  54. (1989). Regulated espression of Mg 2ยง binding epitope on leukocyte integrin o~ subunits.
  55. Regulated expression and binding of three VLA (B1) integrin receptors on T cells.
  56. (1991). Regulation of adhesion to ICAM-1 by the cytoplasmic domain of LFA-1 integrin beta subunit. Science (Wash.
  57. (1992). Regulation of c~zBt-mediated fibroblast migration on type I collagen by shifts in the concentrations of extracellular Mg 2+ and Ca2+.J. Cell Biol.
  58. (1992). Regulation of the VIA integrin-ligand interactions through the B1 subunit.
  59. Role of 8a and/32 integrins in the adhesion of human CD34 hi stem cells to bone marrow stroma.
  60. (1992). Roles for the integrin VLA-4 and its counter receptor VCAM-1 in myogenesis.
  61. (1988). SPa promotor: an ef~cient and versatile mammalian cDNA expression system composed of simian virus 40 early promotor and the R,-U5 segment of human T cell leukemia virus type I long terminal repeat.
  62. (1987). Specific antigen-Ia activation of transfected human T cells expressing Ti c~B-human T3 receptor complexes.
  63. (1990). Structure of the integrin VLA-4 and its cell-cell and cell-matrix adhesion functions.
  64. (1991). T cell receptor dependent, antigen specific stimulation of a routine T cell done induces a transient VLA protein-mediated binding to extracellular matrix, f Immunol.
  65. (1989). T-cell receptor crosslinking transiently stimulates adhesiveness through LFA-1. Nature (Lond.).
  66. (1990). The activation dependent adhesion of macrophages to laminin involves cytoskeleton anchoring and phosphorylation of the alpha-6 beta-1 integrin.
  67. (1991). The cytoplasmic domain of the integrin lymphocyte function-associated antigen 1 ~ subunit: sites required for binding to intercellular adhesion molecule 1 and the phorbol ester-stimulated phosphorylation site.J.
  68. (1990). The leukocyte integrin LFA-1 reconstituted by cDNA transfection in a nonhematopoietic cell line is functionally active and not transiently regulated. Cell Regul.
  69. (1990). The PMA-induced specific association of LFA-1 and talin in intact cloned T helper cells.
  70. (1987). The VLA protein family. Characterization of five distinct cell surface heterodimers each with a common 130,000 molecular weight 13 subunit.
  71. (1991). The VLA-4/VCAM-1 pathway is involved in lymphocyte adhesion to endothelium in rheumatoid synovium.
  72. (1992). to extraceUular matrix and endothelial cells can be regulated through the 13 subunit of VLA.
  73. (1991). Upregulated expression and function of VLA-4 fibronectin receptors on human activated T cells in rheumatoid arthritis.
  74. (1990). VCAM-1 on activated endothelium interacts with the leukocyte integrin VLA-4 at a site distinct from the VLA-4/fibronectin binding site.
  75. (1986). VLA-3: a novel polypeptide association within the VLA molecular complex: cell distribution and biochemical characterization.

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