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pubmed:abstractText |
Previous studies have shown that the addition of reducing agents to the culture medium of embryonic cell lines stimulates their growth. Moreover, recent studies have shown that the redox state of several transcription factors affects their binding to DNA. In light of these findings, we employed gel mobility shift analysis to examine the effects of oxidation and reduction on the ability of transcription factors to bind cis-regulatory elements located in the FGF-4 gene, which is expressed during early mammalian development. In this study, we demonstrate that both the oxidizing agent diamide and the alkylating agent N-ethylmaleimide inhibit the ability of Oct-1, Oct-3, Sp1, and several Sp1-related nuclear proteins to bind important cis-regulatory elements located in the FGF-4 gene. We also demonstrate that not all transcription factors are affected by oxidation. Specifically, we show that the binding of the transcription factor NF-YA, which binds to a critical CCAAT box, and the binding of a high mobility group (HMG) protein(s), which binds to a critical HMG motif, are not affected by diamide or N-ethylmaleimide. Taken together, our findings and those of others support the hypothesis that the redox state of the cell can regulate gene transcription and, thus, can influence important physiological processes.
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