title: Regulation of the avidity of integrin alpha 4 beta 7 by the beta 7 cytoplasmic domain.
abstract: Integrins are cell-surface heterodimeric receptors with adhesive and transmembrane signaling properties. Their cytoplasmic domains can affect receptor avidity, cytoskeletal association, and post-receptor occupancy events. The alpha 4 beta 7 integrin mediates cell adhesion to Peyer's patch high walled endothelial venules (HEV), VCAM, and CS-1/fibronectin. To determine the role of the beta 7 cytoplasmic domain in these interactions, wild-type and truncated beta 7 subunits were stably expressed in the alpha 4+/beta 1-/beta 7- B cell lymphoma 38C13. The cell line delta 727 lacks the entire beta 7 cytoplasmic domain, delta 753 lacks the 34 C-terminal residues, and LXSN is vector-transfected 38C13. Cells expressing wild-type beta 7 bound Peyer's patch HEV, fibronectin, and immobilized VCAM constitutively and did not require prior activation with phorbol esters. Interestingly, delta 753 displayed no ligand binding activity, while delta 727 was constitutively active for all ligands and displayed greater avidity for fibronectin and Peyer's patch HEV than the wild-type beta 7. beta 7, delta 753, delta 727, and LXSN were also tested for the ability to bind soluble VCAM in the presence of various divalent cations. In the presence of Ca2+, but not Mg2+, delta 727 constitutively bound soluble VCAM, whereas beta 7, delta 753, and LXSN did not. beta 7 and delta 753 could bind soluble VCAM if first activated with Mn2+. The results suggest that: (i) alpha 4 beta 7 can be expressed in a constitutively active state, (ii) the beta 7 cytoplasmic domain regulates the avidity of alpha 4 beta 7, and (iii) 38C13 cell lines expressing wild-type and truncated beta 7 subunits define three stable activation states of alpha 4 beta 7: inactive (delta 753), partially active (beta 7), and fully active (delta 727) receptors.
authors.affiliation: Department of Pathology, Stanford University School of Medicine, California 94305.
authors.forename: D T
authors.initials: DT
authors.lastname: Crowe
authors.forename: H
authors.initials: H
authors.lastname: Chiu
authors.forename: S
authors.initials: S
authors.lastname: Fong
authors.forename: I L
authors.initials: IL
authors.lastname: Weissman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D002448:Cell Adhesion; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D004492:Edetic Acid / Q000494:pharmacology; D005353:Fibronectins / Q000378:metabolism*; D005434:Flow Cytometry; D005822:Genetic Vectors; D016023:Integrins / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D016393:Lymphoma, B-Cell; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010581:Peyer's Patches / Q000378:metabolism*; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D002413:Cations, Divalent; D017931:DNA Primers; D005353:Fibronectins; D016023:Integrins; C071777:integrin alpha4beta7; D004492:Edetic Acid; D013755:Tetradecanoylphorbol Acetate