Disruption of integrin function and induction of tyrosine phosphorylation by the autonomously expressed beta 1 integrin cytoplasmic domain.
The cytoplasmic domains of integrin beta subunits are essential for the function of integrins in cell adhesion and signaling. A chimera combining the transmembrane and cytoplasmic domains of the beta 1 integrin subunit with an irrelevant extracellular domain derived from L3T4 (murine CD4) was tested for its ability to interfere with integrin function. Expression of this construct in cultured human embryonic kidney cells under the control of the inducible metallothionein promoter resulted in cell rounding and detachment, and blocked cell adhesion mediated by the beta 1 and alpha v beta 5 integrins. Expression of the beta 1 chimera at basal levels interfered with the tyrosine phosphorylation of a 125-kDa protein induced by antibody-induced clustering of integrins. Induced expression of the chimera resulted in sustained tyrosine phosphorylation of this protein, which could be enhanced by clustering of the chimera but was insensitive to clustering of integrins. These results demonstrate that the autonomously expressed beta 1 integrin cytoplasmic domain can act as a trans-dominant inhibitor of integrin function, presumably via competitive interactions with cytoplasmic components that are required for integrin-mediated cell adhesion and tyrosine phosphorylation.
Lukashev
M E
ME
Department of Medicine, University of California, San Francisco 94143.
Sheppard
D
D
Pytela
R
R
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1; D016023:Integrins / Q000096:biosynthesis / Q000502:physiology*; D007668:Kidney; D007700:Kinetics; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D019000:Phosphotyrosine; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism*; D015183:Restriction Mapping; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D016326:Extracellular Matrix Proteins; D019012:Integrin beta1; D016023:Integrins; D011993:Recombinant Fusion Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D008668:Metallothionein