Determinants contributing to estrogen-regulated expression of SK3.

The rSK3 gene encodes a small conductance Ca(2+)-activated K(+) channel that is transcriptionally regulated by estrogen. To examine determinants of rSK3 gene expression, the CAP site was defined and the promoter was identified. A 33 bp sequence adjacent to the promoter was shown to act as an enhancer in L6 cells that express SK3 and estrogen receptor alpha (ER alpha). The 33 bp enhancer was unable to stimulate transcription in Cos7 cells that do not express SK3 or ER alpha. However when cotransfected with ER alpha and stimulated with 17 beta-estradiol (E2) the enhancer was activated. Interestingly, expression of ER alpha in Cos7 cells and E2 treatment was sufficient to induce expression of the endogenous SK3 gene. Only Sp1 and Sp3 specifically bound to the enhancer from Cos7 and L6 nuclear extracts, however, ER alpha increased Sp1s affinity for the enhancer. The data suggest that estrogen regulates SK3 gene expression through interactions between ER alpha and Sp1.