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In  Drosophila , CLOCK/CYCLE heterodimer (CLK/CYC) is the primary activator of circadian clock genes that contain the E-box sequence in their promoter regions (hereafter referred to as “E-box clock genes”). Although extensive studies have investigated the feedback regulation of clock genes, little is known regarding other factors acting with CLK/CYC. Here we show that Drosophila C-terminal binding protein (dCtBP), a transcriptional co-factor, is involved in the regulation of the E-box clock genes.  In vivo  overexpression of dCtBP in clock cells lengthened or abolished circadian locomotor rhythm with up-regulation of a subset of the E-box clock genes,  period  ( per ),  vrille  ( vri ), and  PAR domain protein 1ε  ( Pdp1ε ). Co-expression of dCtBP with CLK  in vitro  also increased the promoter activity of  per ,  vri ,  Pdp1ε  and  cwo  depending on the amount of dCtBP expression, whereas no effect was observed without CLK. The activation of these clock genes  in vitro  was not observed when we used mutated dCtBP which carries amino acid substitutions in NAD +  domain. These results suggest that dCtBP generally acts as a putative co-activator of CLK/CYC through the E-box sequence.

C-Terminal Binding Protein (CtBP) Activates the Expression of E-Box Clock Genes with CLOCK/CYCLE in Drosophila