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Complementation of dominant suppression implicates CD98 in integrin activation

Abstract

The integrin family of adhesion receptors are involved in cell growth, migration and tumour metastasis1. Integrins are heterodimeric proteins composed of an α and a β subunit, each with a large extracellular, a single transmembrane, and a short cytoplasmic domain. The dynamic regulation of integrin affinity for ligands in response to cellular signals is central to integrin function2. This process is energy dependent and is mediated through integrin cytoplasmic domains3. However, the cellular machinery regulating integrin affinity remains poorly understood. Here we describe a genetic strategy to disentangle integrin signalling pathways. Dominant suppression occurs when overexpression of isolated integrin β1 cytoplasmic domains blocks integrin activation. Proteins involved in integrin signalling were identified by their capacity to complement dominant suppression in an expression cloning scheme. CD98, an early T-cell activation antigen that associates with functional integrins4, was found to regulate integrin activation. Furthermore, antibody-mediated crosslinking of CD98 stimulated β1 integrin-dependent cell adhesion. These data indicate that CD98 is involved in regulating integrin affinity, and validate an unbiased genetic approach to analysing integrin signalling pathways.

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Figure 1: Complementation of dominant suppression and its effect on ligand binding.
Figure 2: Cytoplasmic domain specificity of complementation of dominant suppression.
Figure 3: Human CD98 complements dominant suppression, whereas several other membrane proteins do not.
Figure 4: The affect of a 4F2 monoclonal antibody on adhesion of the small cell lung-cancer cell line (SCLC) H345 to extracellular matrix.

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Acknowledgements

We thank L. Baker for generation of the αβPy cell line and for discussions; S. Shattil and M. Schwartz for suggestions, and reviews of the manuscript; and our colleagues for the reagents acknowledged in the text. This work was supported by grants from the NIH and Cor Therapeutics. T.S. is an MRC (UK) travelling fellow, and P.E.H. is a fellow of the Leukemia Society of America.

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Correspondence to Mark H. Ginsberg.

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Fenczik, C., Sethi, T., Ramos, J. et al. Complementation of dominant suppression implicates CD98 in integrin activation. Nature 390, 81–85 (1997). https://doi.org/10.1038/36349

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