Induction of mouse beta integrin expression following transfection with human alpha 4 chain.
We report here an analysis of the expression and function of the alpha chain of human VLA-4 in stable mouse L cell transfectants and the requirement for the beta chain in these processes. L cells were transfected with human alpha 4 cDNA or alpha 4 and human beta 1 cDNA. Unexpectedly, human alpha 4 cDNA, when transfected alone, could induce de novo surface expression of host beta 7 and increased expression of host beta 1. Induction of mouse beta 7 and beta 1 surface expression was not due to de novo gene activation, but instead represented alpha 4/beta intracellular subunit association and transport to the cell surface. Transfection with human beta 1 prevented surface expression of mouse beta integrins. Whereas human alpha 4 and human beta 1 subunits associated very tightly in anti-alpha 4 immunoprecipitates, human alpha 4 and mouse beta subunits were only partially associated. Furthermore, binding of human/mouse chimeric receptors to recombinant VCAM, a major ligand for alpha 4 beta 7 and alpha 4 beta 1, was very poor, whereas human alpha 4/human beta 1 receptors bound strongly to VCAM. One alpha 4 transfectant, which exhibited a tight human alpha 4/mouse beta 1 association, could be induced, but only after PMA activation, to bind strongly to VCAM. These results indicate that alpha 4 subunits have specific affinity for beta 7 and beta 1 integrins and require beta subunits for surface expression as well as high affinity ligand binding activity. Our results indicate that a tight association between the alpha 4 and beta subunit appears to be critical for ligand binding, consistent with a direct as well as regulatory role for the beta subunit in ligand binding. Furthermore, these studies demonstrate that expression of foreign recombinant proteins can alter host cell protein expression resulting in de novo surface protein expression.
Webb
D L
DL
Institute for Bone and Joint Disorders and Cancer, Bayer Research Center, West Haven, Connecticut 06516, USA.
Conrad
P J
PJ
Ma
L
L
Blue
M L
ML
D000818:Animals; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D039041:Integrin alpha4beta1; D039641:Integrin beta Chains; D019012:Integrin beta1 / Q000096:biosynthesis* / Q000235:genetics; D016023:Integrins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D016201:Receptors, Lymphocyte Homing / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D039041:Integrin alpha4beta1; D039641:Integrin beta Chains; D019012:Integrin beta1; D016023:Integrins; D012333:RNA, Messenger; D016201:Receptors, Lymphocyte Homing; C091368:integrin beta7; D013755:Tetradecanoylphorbol Acetate