Advertisement

Role of the integrin alpha v beta 6 in cell attachment to fibronectin. Heterologous expression of intact and secreted forms of the receptor.

Open AccessPublished:March 04, 1994DOI:https://doi.org/10.1016/S0021-9258(17)37465-3
      This paper is only available as a PDF. To read, Please Download here.
      The integrin alpha v beta 6 has been shown to be a fibronectin-binding protein. To determine whether the cytoplasmic and transmembrane domains of alpha v beta 6 are necessary for binding to fibronectin, a truncated, secreted form of the integrin lacking these domains was engineered and expressed in Chinese hamster ovary cells. Fibronectin affinity chromatography demonstrated that the secreted integrin, like its full-length counterpart, was capable of binding fibronectin. Monoclonal antibodies were made to secreted alpha v beta 6 and to beta 6-transfected NIH 3T3 cells. In experiments designed to determine whether alpha v beta 6 can mediate cell attachment to fibronectin, full-length human beta 6 was expressed in Chinese hamster ovary cells and in the human colon carcinoma cell line SW480. beta 6-expressing cells were identified by alpha v beta 6-specific antibodies, and the beta 6-transfectants were used in cell-adhesion assays. In Chinese hamster ovary cells, human beta 6 associated with hamster alpha v but was incapable of mediating cell attachment to fibronectin. However, expression of beta 6 in these cells had the dominant negative effect of decreasing alpha v beta 5-dependent adhesion to vitronectin. In SW480 cells, beta 6 expression conferred the ability to bind to fibronectin even in the presence of inhibitory antibodies against beta 1 integrins. In such cells, fibronectin binding ability could be blocked by an antibody to alpha v integrins. These results constitute the first direct evidence that alpha v beta 6 mediates cell attachment to fibronectin.

      REFERENCES

        • Hynea R.O.
        Cell. 1987; 48: 549-554
        • Hynes H.O.
        Cell. 1992; 69: 11-25
        • McDonald J.A.
        Am. J. Physiol. 1989; 257: L331-L337
        • Ruoslahti E.
        J. Clin. Invest. 1991; 87: 1-5
        • Busk M.
        • Pytela R.
        • Sheppard D.
        J. Biol. Chem. 1992; 267: 5790-5796
        • Sheppard D.
        • Rozzo C.
        • Starr L.
        • Quaranta V.
        • Erie D.J.
        • Pytela R.
        J. Biol. Chem. 1990; 265: 11502-11507
        • Breuss J.M.
        • Gillett N.
        • Lu L.
        • Sheppard D.
        • Pytela R.
        J. Histochem. Cytochem. 1993; 41: 1521-1527
        • Cheresh D.A.
        • Smith J.W.
        • Cooper H.M.
        • Quaranta V.
        Cell. 1989; 57: 59-69
        • Smith J.W.
        • Vestal D.J.
        • Irwin S.V.
        • Burke T.A.
        • Cheresh D.A.
        J. Biol. Chem. 1990; 265: 11008-11013
        • Engvall E.
        • Ruoslahti E.
        Int. J. Cancer. 1977; 20: 1-5
        • Yatohgo T.
        • Izumi M.
        • Kashiwagi H.
        • Hayashi M.
        Cell Struct. Funct. 1988; 13: 281-292
        • Sheppard D.
        • Cohen D.S.
        • Wang A.
        • Busk M.
        J. Biol. Chem. 1992; 267: 17409-17414
        • Bossy B.
        • Reichardt L.F.
        Biochemistry. 1990; 29: 10191-10198
        • Chuntharapai A.
        • Bodary S.
        • Horton M.
        • Kim K.J.
        Exp. Cell Res. 1993; 205: 345-352
        • Wayner E.A.
        • Orlando R.A.
        • Cheresh D.A.
        J. Cell Biol. 1991; 113: 919-929
        • Dittel B.N.
        • McCarthy J.B.
        • Wayner E.A.
        • LeBien T.W.
        Blood. 1993; 81: 2272-2282
        • Brown P.J.
        • Juliano R.L.
        Science. 1985; 228: 1448-1450
        • Brown P.J.
        • Juliano R.L.
        Exp. Cell Res. 1988; 171: 303-318
        • Houghton A.N.
        • Eisenger M.
        • Albino A.P.
        • Caimcross J.G.
        • Old L.J.
        J. Exp. Med. 1982; 156: 1755-1766
        • Real F.X.
        • Houghton A.N.
        • Albino A.P.
        • Cordon-Cardo C.
        • Melamed M.R.
        • Oettgen H.F.
        • Old L.J.
        Cancer Res. 1985; 45: 1411-4401
        • Sambrook J.
        • Fritsch E.F.
        • Maniatis T.
        Molecular Cloning: A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY1989
        • Bodary S.C.
        • McLean J.W.
        J. Biol. Chem. 1990; 265: 5938-5941
        • Briesewitz R.
        • Epstein M.R.
        • Marcantonio E.E.
        J. Biol. Chem. 1993; 268: 2989-2996
        • Dana N.
        • Fathallah D.M.
        • Arnaout M.A.
        Proc. Natl. Acad. Sci. U. S. A. 1991; 88: 3106-3110
        • Frachet P.
        • Duperray A.
        • Delachanal E.
        • Marguerie G.
        Biochemistry. 1992; 31: 2408-2415
        • Hayashi Y.
        • Haimovich B.
        • Reszka A.
        • Boettiger D.
        • Horwitz A.
        J. Cell Biol. 1990; 110: 175-184
        • Solowska J.
        • Guan J.L.
        • Marcantonio E.E.
        • Trevithick J.E.
        • Buck C.A.
        • Hynes R.O.
        J. Cell Biol. 1989; 109: 853-861
        • Solowska J.
        • Edelman J.M.
        • Albelda S.M.
        • Buck C.A.
        J. Cell Biol. 1991; 114: 1079-1088
        • Zhang Z.
        • Morla A.O.
        • Vuori K.
        • Bauer J.S.
        • Juliano R.L.
        • Ruoslahti E.
        J. Cell Biol. 1993; 122: 235-242
        • Takada Y.
        • Ylanne J.
        • Mandelman D.
        • Puzon W.
        • Ginsberg M.H.
        J. Cell Biol. 1992; 119: 913-921