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Background  The Notch signaling pathway is fundamental to the regulation of many cell fate decisions in eumetazoans. Not surprisingly, members of this pathway are highly conserved even between vertebrates and invertebrates. There is one notable exception, Hairless, which acts as a general Notch antagonist in  Drosophila . Hairless silences Notch target genes by assembling a repressor complex together with Suppressor of Hairless [Su(H)] and the co-repressors Groucho (Gro) and C-terminal binding protein (CtBP). Now with the availability of genomic databases, presumptive Hairless homologues are predicted, however only in insect species. To further our understanding of Hairless structure and function, we have cloned the  Hairless  gene from  Apis mellifera  ( A.m.H ) and characterized its functional conservation in  Drosophila .    Results  The  Apis  Hairless protein is only one third of the size of the  Drosophila  orthologue. Interestingly, the defined Suppressor of Hairless binding domain is interrupted by a nonconserved spacer sequence and the N-terminal motif is sufficient for binding. In contrast to  Apis  Hairless, the  Drosophila  orthologue contains a large acidic domain and we provide experimental evidence that this acidic domain is necessary to silence Hairless activity in vivo. Despite the dramatic size differences,  Apis  Hairless binds to the  Drosophila  Hairless interactors Su(H), Gro, CtBP and Pros26.4. Hence,  Apis  Hairless assembles a repressor complex with  Drosophila  components that may have a different topology. Nevertheless,  Apis  Hairless is sufficient to repress the Notch target gene  vestigial  in  Drosophila . Moreover, it is able to rescue  Hairless  mutant phenotypes, providing in vivo evidence for its function as a bona fide Notch antagonist.    Conclusion  This is the first interspecies-complementation analysis of the Hairless gene. Guided by evolutionary comparisons, we hope to eventually identify all the relevant structural domains and cofactors of Hairless, thereby opening an avenue for further insights into the repressor-complexes that down-regulate Notch signaling also in other, higher eukaryotes.

The tiny Hairless protein from Apis mellifera: a potent antagonist of Notch signaling in Drosophila melanogaster