5-AzaC Treatment Enhances Expression of Transforming Growth Factor-β Receptors through Down-regulation of Sp3

Sudhakar Ammanamanchi, Michael G Brattain

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

We have previously reported that Sp3 acts as a transcriptional repressor of transforming growth factor-β receptors type I (RI) and type II (RII). We now present data suggesting that treatment of MCF-7L breast and GEO colon cancer cells with 5-aza cytidine (5-azaC) leads to down-regulation of Sp3 and the concomitant induction of RI and RII. Western blot and gel shift analyses on 5-azaC-treated MCF-7L and GEO nuclear extracts indicated reduced Sp3 protein levels and decreased binding of Sp3 protein to radiolabeled consensus Sp1 oligonucleotide. Southwestern analysis detected decreased binding of Sp3 to RI and RII promoters in 5-azaC-treated MCF-7L and GEO cells, suggesting a correlation between decreased Sp3 binding and enhanced RI and RII expression in these cells. Reverse transcription-polymerase chain reaction and nuclear run-on data from 5-azaC-treated MCF-7L and GEO cells indicated down-regulation of Sp3 mRNA as a result of decreased transcription of Sp3. We reported earlier that 5-azaC treatment induces RI and RII expression through increased Sp1 protein levels/activities in these cells. These studies demonstrate that the effect of 5-azaC involves a combination of effects on Sp1 and Sp3.

Original languageEnglish (US)
Pages (from-to)32854-32859
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number35
DOIs
StatePublished - Aug 31 2001

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Cytidine
Growth Factor Receptors
Transforming Growth Factors
Down-Regulation
Transcription
Proteins
Polymerase chain reaction
Electrophoretic Mobility Shift Assay
Oligonucleotides
Colonic Neoplasms
Reverse Transcription
Carrier Proteins
Breast
Western Blotting
Gels
Cells
Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

5-AzaC Treatment Enhances Expression of Transforming Growth Factor-β Receptors through Down-regulation of Sp3. / Ammanamanchi, Sudhakar; Brattain, Michael G.

In: Journal of Biological Chemistry, Vol. 276, No. 35, 31.08.2001, p. 32854-32859.

Research output: Contribution to journalArticle

Ammanamanchi, Sudhakar ; Brattain, Michael G. / 5-AzaC Treatment Enhances Expression of Transforming Growth Factor-β Receptors through Down-regulation of Sp3. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 35. pp. 32854-32859.
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