MULTISTEP REGULATION OF ENHANCER ACTIVITY OF THE 21-BASE-PAIR ELEMENT OF HUMAN T-CELL LEUKEMIA-VIRUS TYPE-I

We examined the regulatory mechanisms of binding and transcriptional enhancement of the 21-bp core element of the enhancer of human T-cell leukemia virus type I (HTLV-1) in response to forskolin, 12-O-tetradecanoylphorbol-13-acetate (TPA), and a viral transactivator, p40tax. The 21-bp core element has been shown to bind to a cyclic AMP-responsive element binding protein (CREB)-like molecule at the site of an imperfect palindrome containing the TGAC motif. Experiments with oligonucleotides with mutations in the imperfect palindrome demonstrated that one TGAC motif is necessary and sufficient for both the binding of the CREB-related factor and transcriptional activity in response to forskolin in a human T-cell line, Jurkat. We also found that binding of the CREB-like factor to the 21-bp core element was enhanced by treatment with TPA, with little effect on transcriptional activity; in contrast, forskolin and p40tax did not facilitate binding, though they enhanced transcription. The combination of forskolin and TPA synergistically induced the transcription activity of the element, showing a hierarchical mechanism of regulation of the HTLV-I core enhancer element to levels sufficient for formation of the factor-enhancer complex and for activation of the complex. Added to those findings, our results indicate that the modes of activation by forskolin and p40tax are different from each other.