Functional Characterization of Transforming Growth Factor β Type II Receptor Mutants in Human Cancer

We recently identified missense mutations at amino acid residues 526 and 537 located within the highly conserved subdomain XI of the transforming growth factor β type II receptor (TβR-II) serine-threonine kinase in two human squamous carcinoma cell lines. These cell lines are resistant to transforming growth factor β-mediated inhibition of growth. Moreover, treatment with transforming growth factor β fails to increase the levels of type 1 plasminogen activator inhibitor and fibronectin synthesis. To test the effects of the mutations on receptor function, mutant TβR-II cDNAs were expressed in TβR-II-deficient T47D cells. Cyclin A promoter activity was reduced by 50% in cells expressing wild-type TβR-II but increased 2-fold in cells transfected with either of the two mutant receptors. Conversely, plasminogen activator inhibitor type 1 promoter activity was increased 6-fold in cells transfected with wild-type receptor but not with either of the two mutant receptors. Moreover, the activity of both mutant serine-threonine kinases was strongly reduced compared to that of the wild-type receptor. Thus, the amino acid residues at positions 526 and 537 seem to be essential for kinase function and signaling activity of the TβR-II.

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