The dorsal protein enhances the biosynthesis and stability of the Drosophila I kappa B homologue cactus

Abstract

The cactus and dorsal proteins are Drosophila homologues of mammalian I kappa B cytoplasmic anchor proteins and rel/NF kappa B transcription factors respectively. They are required for the generation of embryonic dorsoventral polarity and probably at later developmental stages for an innate immune response. In this paper we report on the properties of SLDL, a derivative of the SL2 cell line in which dorsal is expressed constitutively. In SLDL cells biosynthesis of cactus protein is stimulated by approximately 4-fold when compared with SL2 cells. Enhanced biosynthesis of cactus protein cannot be explained solely on the basis of increased expression of the cactus gene as the level of the corresponding mRNA is only 2-fold higher than in SL2 cells. On the basis of these findings we propose that free cytoplasmic dorsal protein is able, directly or indirectly to stimulate translation of the cactus mRNA. Such an arrangement would enable the dorsal protein to be buffered in the cytoplasm of the resting cell over a wide range of concentrations. We also show here that subsequent to biosynthesis the cactus protein is either rapidly degraded or incorporated into complexes with dorsal. Protein that does not associate with dorsal has a half-life of approximately 40 min whereas that which is incorporated into complexes is very stable, having a half life in excess of 24 h. The complexed cactus protein is acted on by protein kinases which generate distinct phophorylated isoforms.