Several Notch interactions occur in rapid succession during early C.elegans embryogenesis, each resulting in a distinct fate. We have previously shown that
ref-1, a gene distantly related to Drosophila E(spl), is a direct Notch target in all these interactions. We show here that
ref-1 expression is controlled by multiple, Notch-dependent enhancers that are specific for each interaction. We have identified a 150bp enhancer that is specific to Notch interactions in the endoderm, and found similar enhancers with the same activity in C.briggsae and C.remanei. The endoderm enhancer contains multiple, conserved binding sites for LAG-1/Su(H) and GATA transcription factors. We demonstrate that all LAG-1/Su(H) and GATA sites are required for full activity of this enhancer. We provide evidence that a GATA transcription factor called ELT-2 is a cooperative factor for Notch in the endoderm. Ectopic expression of ELT-2 in non-endodermal lineages caused activation of the endoderm enhancer only in Notch-signaled cells, suggesting that the presence of the cooperative factor dictates in which Notch interaction a Notch-dependent enhancer becomes responsive in vivo. Previous studies in Drosophila showed that synergy between Su(H) and the bHLH transcription factor Daughterless requires a specific configuration of Su(H) sites, known as a Su(H) paired site, SPS. The endoderm enhancer contains a putative SPS. However, we find that Notch-GATA synergy does not require a SPS, and instead requires a non-SPS configuration of oriented LAG-1/Su(H) sites. Thus, it appears that that each configuration couples Notch signaling with a specific class of transcription factor.