NR2E3 transcriptional repression activity requires the formation of a functional dimer.

Abstract

<p><b>A</b>. Top view of the NR2E3 LBD dimer, showing the close interaction of L372 and L375 (red and orange stick models, resp.) from the helices 10 (cyan) in the dimer interface. <b>B</b>. A close-up view of the helices 10 in the dimer interface. <b>C</b>. Size Exclusion Chromatography for Bio-rad Protein Standard (left) and MBP-NR2E3 LBD (right). <b>D</b>. Mutation of helix 10 coiled coil interface residues abolished LBD dimerization in a mammalian two hybrid assay. Reporter gene activation by Gal4DBD–NR2E3LBD and VP16AD–NR2E3LBD wildtype and mutant expression plasmids is shown as bar graph. Cells cotransfected with pBIND-Id and pACT-MyoD were used as positive controls. <b>E</b>. Effects of the mutations L372R, L375R, and the double mutation L372R/L375R on NR2E3 repression activity (top). Below: Expression levels of wildtype and mutant Gal4DBD–NR2E3LBD determined by immunoblotting.</p