Direct homeodomain–DNA interaction in the autoregulation of the fushi tarazu gene

Abstract

A MAJOR problem in the elucidation of the molecular mechanisms governing development is the distinction between direct and indirect regulatory interactions among developmental control genes^1–5. In vivo studies have indicated that the Drosophila segmentation genefushi tarazu (ftz) directly or indirectly autoregulates its expression⁶. Here we describe a generally applicable experimental approach which establishes a direct in vivo interaction of the homeodomain protein ftz with the ftz cis-autoregulatory control region.In vitro studies have shown that the DNA-binding specificity of the ftz homeodomain can be changed by a single amino-acid substitution in the recognition helix (Gin 50 →"Lys)⁷. Whereas wild-type ftz homeodomain binds preferentially to a CCATTA motif, the mutant homeodomain (ftzQSOK) recognizes a GG ATT A motif. We now find that the in vivo activity of an ftz autoregulatory enhancer element is reduced by mutations of putative ftz-binding sites to GGATTA. This down-regulatory effect is specifically suppressed in vivo by the DNA-binding specificity mutantftzQ5OK. These results establish a direct positive autoregulatory feedback mechanism in the regulation of this homeobox gene.