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Papers In Press, published online ahead of print October 10, 2000
J. Biol. Chem, 10.1074/jbc.M002462200
Submitted on March 22, 2000
Revised on October 10, 2000
Accepted on October 10, 2000
Department of Surgery, The University of Texas, San Antonio, TX 78284
Corresponding Author: brattainm{at}uthscsa.edu
MCF-7E breast cancer cells express transforming growth factor-b receptors RI and RII in comparison to MCF-7L cells. We present data showing that Sp3 acts as a transcriptional repressor of RI and RII in MCF-7L cells and GEO colon cancer cells. MCF-7L and GEO cells express high levels of Sp3 protein. Gel shift analysis indicated enhanced binding of Sp3 from MCF-7L cells to a consensus Sp1 oligonucleotide. Southwestern data indicated increased binding of Sp3 to RI and RII promoters in MCF-7L cells suggesting a correlation between Sp3 binding and reduced expression of TGF-b receptors in MCF-7L cells. Co-transfection of CMV-Sp3 cDNA with RI and RII promoter-luciferase reporter constructs decreased RI and RII promoter activities by 70% in MCF-7E and GEO cells. Southwestern analysis detected the binding of transiently expressed Sp3 to RI and RII promoters in MCF-7E cells. Significantly, ectopic Sp3 expression led to repression of RI and RII transcripts in MCF-7E cells. This report demonstrates that inappropriate over expression of Sp3 is a mechanism which contributes to repression of TGF-b receptors.
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