cis-regulatory analysis of cat-2/tyrosine hydroxylase effector gene reveals functional binding sites for UNC-62 and VAB-3. (A) Multicopy extrachromosomal reporter analysis of cat-2 minimal dopaminergic cis-regulatory module (cat-2p21). In addition to published functional AST-1/ETS, CEH-20/PBX HD, CEH-43/DLL HD, binding sites (Flames and Hobert 2009; Doitsidou et al. 2013), UNC-62/MEIS and VAB-3/PAIRED binding sites are also required for correct GFP reporter expression in different dopaminergic neuron types. HD* represents a PAIRED-type HD consensus (HTAATTR). Black crosses represent point mutations to disrupt the corresponding TFBS, the nature of the mutation is indicated in red. (+) >70% of mean wild-type construct values; (−) values are less than 10% expression in at least two of the three analyzed lines. n > 30 animals per line. (B) Analysis of the effect of MEIS and PAIRED TFBS cis mutations in the expression of endogenous cat-2 gene. Schema of used strains with the corresponding mutations, quantification of reporter gene expression defects, and representative micrographs are shown. n > 50 animals each condition; (*) P < 0.05 compared to wild type. Scale: 25 µm.
