Cloning and characterization of the rat Hsf2 promoter: a critical role of proximal E-box element and USF protein in Hsf2 regulation in different compartments of the brain.
The complex patterns of tissue-, cell type- and developmental stage-specific expression of heat shock factor 2 (Hsf2) raise a question of how this can be achieved for this ubiquitous transcription factor. To explore molecular mechanisms responsible for the regulated expression of Hsf2, a 2638-bp 5'-flanking region of the rat Hsf2 gene was cloned and characterized. Since the brain represents one of the most complicated organs composed of several regions with different cell types, differential regulation of Hsf2 in various brain regions was investigated in detail. Results show that the major transcription initiation site of the Hsf2 gene is located at cytosine-155 relative to the translation initiation site. The E-box element located immediate upstream of the transcription initiation site was demonstrated to be critical for Hsf2 promoter activity, and the upstream stimulatory factor (USF) protein was identified as the major E-box binding protein. That the only two base exchange of the E-box core sequences from CACGTG to CACGGT severely impaired Hsf2 promoter activity and completely eliminated USF binding clearly demonstrated that the specific binding of USF to E-box is critical for Hsf2 promoter activity. Here we demonstrated that the Hsf2 expression levels varied significantly in different brain regions. We also demonstrated that Hsf2 expression levels in various brain regions relatively correlated with the E-box binding activity of USF. Based on these results, we suggest that E-box binding activity of USF protein may act as one of the major regulators of Hsf2 expression in situ although a possible involvement of other transcription factors cannot be ruled out. The presence of several transcription factor binding sites of biological importance in the Hsf2 promoter suggests that identifying the interplay of USF and these factors should help further elucidate the molecular mechanisms of tissue-, cell type- and developmental stage-specific expression of Hsf2.[1]References
- Cloning and characterization of the rat Hsf2 promoter: a critical role of proximal E-box element and USF protein in Hsf2 regulation in different compartments of the brain. Lee, S.S., Kwon, S.H., Sung, J.S., Han, M.Y., Park, Y.M. Biochim. Biophys. Acta (2003) [Pubmed]
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