Abstract
The protein synthesis inhibitor cycloheximide (Chx) suppresses prolactin-induced β-casein gene expression in the mammary epithelial cell line COMMA-D. As the mechanism underlying this effect is unclear, the effects of protein synthesis inhibitors on interactions of transcription factors with the β-casein promoter were examined. Suppression of prolactin-induced β-casein gene expression occurred in both COMMA-D cells and primary mammary cell cultures with as little as 2 h protein synthesis inhibition. This was associated with changes in transcription factors interacting at a response element in the proximal region of the rat β-casein promoter. Inhibition of protein synthesis was associated with NF-κB binding at a site immediately 3' to the Stat5-binding site at position 97–89 of the β-casein promoter, suppression of Stat5 DNA-binding activity, and inhibition of Stat5 tyrosine phosphorylation. Treatment with the NF-κB inhibitor parthenolide failed to restore prolactin responsiveness. These results show that protein synthesis inhibition is associated with both blockage of prolactin-Stat5 signaling and NF-κB binding to the β-casein promoter, but that the latter is not necessary for the suppression of β-casein expression.
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Beaton, A., Broadhurst, M.K., Wilkins, R.J. et al. Suppression of β-casein gene expression by inhibition of protein synthesis in mouse mammary epithelial cells is associated with stimulation of NF-κB activity and blockage of prolactin-Stat5 signaling. Cell Tissue Res 311, 207–215 (2003). https://doi.org/10.1007/s00441-002-0672-2
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DOI: https://doi.org/10.1007/s00441-002-0672-2