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Mechanism of Induction of Transforming Growth Factor-ß Type II Receptor Gene Expression by v-Src in Murine Myeloid Cells

Laboratory of Cell Regulation and Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892-5055 [S. H. P., Y. Y., B. I. L., D. K. L., S-J. K.], and Intramural Research Support Program, Science Applications International Corporation-Frederick [M. C. B-R., T. F.], Laboratory of Leukocyte Biology, Frederick Cancer Research and Development Center [D. C. B., F. R.], Frederick, Maryland 21702

Abstract

Transforming growth factor (TGF)-ß1 plays an important role during hematopoiesis. Previously, we had shown that the growth of a v-Src-transformed myeloid cell line was markedly more inhibited by TGF-ß treatment when compared with the wild-type myeloid cell line. To investigate the increased growth sensitivity of the v-Src-transformed myeloid cell line, 32D-src, to TGF-ß, we examined expression of the TGF-ß type II receptor (TGF-ß RII) gene in myeloid cell lines. Northern blot analysis showed that expression of 8- and 6-kb species of TGF-ß RII transcripts was markedly increased in the 32D-src cell line. The expression of the TGF-ß RII promoter linked to a reporter gene was increased 23-fold by v-Src. DNA transfection and electrophoretic mobility shift assay revealed that v-Src induces TGF-ß RII promoter activity through an AP1/ATF2-like sequence (-219 to -172), ETS binding sites (+1 to +36), and the inverted CCAAT box (-81 to -77). Novel DNA-protein complexes with ETS binding sites are significantly increased in v-src-transformed cell lines compared with the control cell line. These results suggest that v-Src induces activity of the TGF-ß RII promoter through multiple elements by inducing expression of nuclear proteins interacting with these elements.

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