Proteins associated with RORalpha binding element on SULT1E1 gene

Estrogen sulfotransferase (SULT1E1) metabolically inactivates estrogen and SULT1E1 expression is tightly regulated by multiple nuclear receptors in cells. A series of biochemical techniques have clearly demonstrated that nuclear receptors HNF4alpha and phosphorylated RORalpha at Ser100 form as complexes binding to the SULT1E1 enhancer to activate gene transcription in response to a high glucose (450 mg/dL) signal. However, the kinase that mediates this phosphorylation at high glucose condition remain in mysterious. To identify these binding factors and possible kinases that mediate the phosphorylation on the SULT1E1 enhancer, the SULT1E1 enhancer was biotinylated and conjugated to magnetic beads, which were subsequently used to enrich the binding factors from low glucose (40 mg/dL) and high glucose (450 mg/dL) treated HepG2 cell nuclear extracts.