engrailed (en) is a gene important for many different aspects of development including embryonic segmentation and formation of the posterior compartment in imaginal discs. en and the co-regulated, redundant gene inv reside in a 113 kb domain covered with tri-methylated histone H3 (H3K27me3), the distinctive mark of genes regulated by the Polycomb group proteins. We have been identifying en enhancers that drive its complex expression during development. We previously reported that a 79kb genomic en construct (HA-en79) contains enhancers for both embryonic and wing imaginal disc (WD) expression, and rescues a chromosomal deletion of the inv/en domain. Interestingly, even though the WD enhancers are present in the transgene, their ability to drive HA-en expression is subject to en protein (EN) auto-repression; in the presence of endogenous EN, HA-en is only expressed in subsets of the wild type domain. In contrast, the endogenous en gene is not repressed by the presence of the HA-en79 transgene. Further, reporter constructs that contain the imaginal disc enhancers driving lacZ show lacZ expression only in the anterior compartment; our data strongly suggest that EN silences lacZ expression in the posterior compartment. These data indicate that removing the imaginal disc enhancers from the inv/en domain alters their activity, rendering them sensitive to repression by EN. Chromatin-immunoprecipitation experiments suggest that EN binds directly to a DNA fragment that contains one of the imaginal disc enhancers. We suggest that when this imaginal disc enhancer is within the inv/en domain, its activity is high, and it cannot be repressed by EN, but when this same enhancer is taken out of the domain, EN is able to repress it. Experiments to examine the chromatin marks present in the endogenous locus and the small and large transgene are in the planning stages.