Erica M. Selva

Sprinter: a novel transmembrane protein required for Wg secretion and signaling.

Wingless (Wg) is a secreted ligand that differentially activates gene expression in target tissues. It belongs to the Wnt family of secreted signaling molecules that regulate cell-to-cell interactions during development. Activation of Wg targets is dependent on the ligand concentration in the extracellular milieu; cellular mechanisms that govern the synthesis, delivery and receipt of Wg are elaborate and complex. We have identified sprinter (srt), which encodes a novel, evolutionarily conserved transmembrane protein required for the transmission of the Wg signal. Mutations in srt cause the accumulation of Wg in cells that express it, and retention of the ligand prevents activation of its target genes in signal-receiving cells. In the absence of Srt activity, levels of Wg targets (including Engrailed in embryos lacking maternal and zygotic srt, and Senseless and Achaete in wing discs) are reduced. Activation of Wg targets in the receiving cells does not require srt. Hence, the function of Srt is restricted to events occurring within the Wg-producing cells. We show that srt is not required for any aspect of Hedgehog (Hh) signal transduction, suggesting specificity of srt for the Wg pathway. We propose that srt encodes a protein required for Wg secretion that regulates maturation, membrane targeting or delivery of Wg. Loss of srt function in turn diminishes Wg-pathway activation in receiving cells.

Dual role of the fringe connection gene in both heparan sulphate and fringe-dependent signalling events

The precise regulation of growth factor signalling is crucial to the molecular control of development in Drosophila. Post-translational modification of signalling molecules is one of the mechanisms that modulate developmental signalling specificity. We describe a new gene, fringe connection (frc), that encodes a nucleotide–sugar transporter that transfers UDP–glucuronic acid, UDP–N-acetylglucosamine and possibly UDP–xylose from the cytoplasm into the lumen of the endoplasmic reticulum/Golgi. Embryos with the frc mutation display defects in Wingless, Hedgehog and fibroblast growth factor signalling. Clonal analysis shows that fringe-dependent Notch signalling is disrupted in frc mutant tissue.

slalom encodes an adenosine 3′‐phosphate 5′‐phosphosulfate transporter essential for development in Drosophila

Sulfation of all macromolecules entering the secretory pathway in higher organisms occurs in the Golgi and requires the high‐energy sulfate donor adenosine 3′‐phosphate 5′‐phosphosulfate. Here we report the first molecular identification of a gene that encodes a transmembrane protein required to transport adenosine 3′‐phosphate 5′‐phosphosulfate from the cytosol into the Golgi lumen. Mutations in this gene, which we call slalom, display defects in Wg and Hh signaling, which are likely due to the lack of sulfation of glycos aminoglycans by the sulfotransferase sulfateless. Analysis of mosaic mutant ovaries shows that sll function is also essential for dorsal–ventral axis determination, suggesting that sll transports the sulfate donor required for sulfotransferase activity of the dorsal–ventral determinant pipe.

Differential effects of the mismatch repair genes MSH2 and MSH3 on homeologous recombination in Saccharomyces cerevisiae.

Abstract The products of the yeast mismatch repair genes MSH2 and MSH3 participate in the inhibition of genetic recombination between homeologous (divergent) DNA sequences. In strains deficient for these genes, homeologous recombination rates between repeated elements are elevated due to the loss of this inhibition. In this study, the effects of these mutations were further analyzed by quantitation of mitotic homeologous recombinants as crossovers, gene conversions or exceptional events in wild-type, msh2, msh3 and msh2 msh3 mutant strains. When homeologous sequences were present as a direct repeat in one orientation, crossovers and gene conversions were elevated in msh2, msh3 and msh2 msh3 strains. The increases were greater in the msh2 msh3 double mutant than in either single mutant. When the order of the homeologous sequences was reversed, the msh2 mutation again yielded increased rates of crossovers and gene conversions. However, in an msh3 strain, gene conversions occurred at higher levels but interchromosomal crossovers were not increased and intrachromosomal crossovers were reduced relative to wild type. The msh2 msh3 double mutant behaved like the msh2 single mutant in this orientation. Control strains harboring homologous duplications were largely but not entirely unaffected in mutant strains, suggesting specificity for the mismatched intermediates of homeologous recombination. In all strains, very few (<10%) recombinants could be attributed to exceptional events. These results suggest that MSH2 and MSH3 can function differentially to control homeologous exchanges.

Role of heparan sulfate proteoglycans in cell signaling and cancer

Publisher Summary Heparan sulfate proteoglycans (HSPGs) are composed of a protein core modified on specific serine residues by the addition of heparan sulfate (HS) glucosaminoglycans (GAGs) synthesized in the Golgi. Three distinct types of proteins can serve as the HSPG cores: the transmembrane proteins encoded by the syndecan genes, the glycosylphosphatidylinositol (GPI) membrane-bound glypicans, and the extracellular matrix secreted perlecan proteins. The chapter presents the two major advances made in understanding the role of HSPGs in development and cancer. First, studies in Drosophila have identified many mutations in either the biosynthetic enzymes for example, Sugarless (Sgl), Sulfateless (Sfl), Tout velu (Ttv); or the protein cores for example, syndecans and glypicans. Analysis of the mutant phenotypes has revealed the critical roles of HSPGs in modulating various growth factor signaling pathways. Second, many mutations linked to human cancers have been isolated and shown to correspond to defects in the biosynthesis of HSPGs. The chapter explains the importance of HSPGs in cell signaling and provides insights into their functions. Modulating the activity of HSPGs could influence the development of cancers caused by aberrant cell signaling. The chapter concludes that HSPGs constitute important targets for therapeutics to treat some human tumors.