Sequence motifs in the integrin alpha 4 cytoplasmic tail required for regulation of in vivo expansion of murine lymphoma cells

Abstract

The binding of integrins to cognate ligands is tightly controlled by intracellular signals. Conversely, integrin occupancy generates biochemical signals inside the cell. The present study examined whether concepts of integrin function established by in vitro analysis apply to regulation of receptor function in complex biologic settings in vivo using a mouse model of tumor metastasis. Integrin alpha 4 subunits were truncated at amino acid Gln1014 (A4-1014), preserving the conserved GFFKR motif, and at position Glu1021 (A4-1021). In vitro adhesion assays revealed that cytoplasmic tail truncations did not affect constitutive ligand binding of alpha 4 integrins, while agonist-induced adhesion was abolished by the alpha 4-1014, but not by the alpha 4-1021, mutation. Inducible ligand binding of alpha 4 integrins was dependent on cytoskeletal function, whereas constitutive adhesion was not. In vivo metastasis formation assays demonstrated that expansion of murine T lymphoma cells in spleen is strongly inhibited by the wild-type alpha 4 subunit and the alpha 4-1021 mutant. In contrast, the in vivo phenotype of alpha 4 integrin expression in lymphoma cells was completely abrogated by the alpha 4-1014 mutation. Cross-linking of alpha 4 integrins in vitro inhibited proliferation and induced apoptosis of LB cells expressing wild-type alpha 4 subunits or the alpha 4-1021 mutant, but not of LB-A4-1014 cells. In summary, these results demonstrate that sequence motifs regulating cytoskeleton-dependent alpha 4 integrin activation in vitro are essential for the control of LB lymphoma cell expansion both in vitro and in vivo.