Genetic analysis shows that Engrailed (En), a homeodomain-containing transcription factor, has both negative and positive targets. Negative regulation is expected from a factor that has a well-defined repressor domain but activation is harder to comprehend. We used VP16En, a form of En that had its repressor domain replaced by the activation domain of VP16, to show that En activates targets using two parallel routes, by repressing a repressor and by being a bona fide activator. We identified the intermediate repressor activity as being encoded by sloppy paired 1 and 2 and showed that bona fide activation is dramatically enhanced by Wingless signaling. Thus, En is a bifunctional transcription factor and the recruitment of additional cofactors presumably specifies which function prevails on an individual promoter. Extradenticle (Exd) is a cofactor thought to be required for activation by Hox proteins. However, in thoracic segments, Exd is required for repression (as well as activation) by En. This is consistent with in vitro results showing that Exd is involved in recognition of positive and negative targets. Moreover, we provide genetic evidence that, in abdominal segments, Ubx and Abd-A, two homeotic proteins not previously thought to participate in the segmentation cascade, are also involved in the repression of target genes by En. We suggest that, like Exd, Ubx and Abd-A could help En recognize target genes or activate the expression of factors that do so.