title: Individual embryonic fibroblasts express multiple beta chains in association with the alpha v integrin subunit. Loss of beta 3 expression with cell confluence.
abstract: The alpha chain of the vitronectin receptor, alpha v, has been found in association with the integrin subunits beta 1, beta 3, or beta 5 on different cell types. We show here that cultured embryonic fibroblasts simultaneously display alpha v beta 3, alpha v beta 1, and alpha v in association with two other beta subunits, one of which is probably beta 5. Polymerase chain reaction analysis of single cells isolated by micromanipulation identified mRNA for alpha v, beta 1, beta 3, and beta 5 in six of eight clones. Immunoprecipitation of iodinated cell surface proteins with a monoclonal antibody to alpha v indicated that the relative proportions of the different beta chains in association with alpha v varied, particularly between two different cell lines. The cytokines platelet-derived growth factor, transforming growth factor beta 1, and tumor necrosis factor alpha did not appear to alter this ratio although tumor necrosis factor alpha increased the surface expression of the alpha v-associated integrins; but overnight culture in basic fibroblast growth factor caused a lower expression of alpha v beta 1 and alpha v beta 5 with no reduction in alpha v beta 3 expression. When the cell cultures were grown to complete confluence, surface expression of beta 3 was abolished, and the expression of an unknown beta chain (beta u) became more prominent. This effect was not overcome by culturing confluent cells with basic fibroblast growth factor. Affinity column chromatography showed that alpha v beta 5 bound to vitronectin but alpha v beta 1 did not, whereas alpha v beta 1 but not alpha v beta 5 bound to fibronectin. These results suggest that, on individual cells, the beta subunits found in association with alpha v may vary according to the proliferative capacity of the cell and that the promiscuous beta 3 subunit is progressively replaced by beta subunits of individual ligand specificity.
authors.affiliation: Cancer Research Unit, Faculty of Medicine, University of Newcastle, New South Wales, Australia.
authors.forename: R C
authors.initials: RC
authors.lastname: Bates
authors.forename: L M
authors.initials: LM
authors.lastname: Rankin
authors.forename: C M
authors.initials: CM
authors.lastname: Lucas
authors.forename: J L
authors.initials: JL
authors.lastname: Scott
authors.forename: G W
authors.initials: GW
authors.lastname: Krissansen
authors.forename: G F
authors.initials: GF
authors.lastname: Burns
mesh_terms: D001483:Base Sequence; D002455:Cell Division; D002478:Cells, Cultured; D003854:Deoxyribonucleotides; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008024:Ligands; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000378:metabolism; D011971:Receptors, Immunologic / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D019030:Receptors, Vitronectin; D019096:Vitronectin
chemical_list: D003854:Deoxyribonucleotides; D005353:Fibronectins; D006023:Glycoproteins; D016023:Integrins; D008024:Ligands; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D019030:Receptors, Vitronectin; D019096:Vitronectin