IBCN   20355
INSTITUTO DE BIOLOGIA CELULAR Y NEUROCIENCIA "PROFESOR EDUARDO DE ROBERTIS"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Functional characterization of SLC35A3 mutants associated with congenital disorders of glycosylation and the use of Xenopus as a biological model.
Autor/es:
LÓPEZ, SILVIA L.; FAVAROLO, MARÍA BELÉN; BREDESTON, LUIS M.
Lugar:
Buenos Aires
Reunión:
Congreso; III International Congress in Translational Medicine; 2017
Resumen:
Nucleotide-sugar transporters (NSTs) are integral membrane proteins of the Golgi apparatus and the Endoplasmic Reticulum (ER), that controls the flux of nucleotide-sugars from the cytosol to the lumen, where they serve as substrate for specific glycosyltransferases during the synthesis of diverse glycoconjugates (glycolipids, glycoproteins and proteoglycans). The NSTs are members of SLC35 family, a group of highly conserved hydrophobic proteins with multiple transmembrane domains with a size of 30-40kDa. An antiporter mechanism, nucleotide sugar/ nucleoside monophosphate, have been proposed (Hirschberg et al 1998). Nevertheless the molecular basis of ligand recognition and transport is unknown. Like other membrane transport proteins the progress in structure-function studies are limited due to the complexity in the transport activity characterization assays and the difficulty to obtain large amounts of purified protein. It was described that mutations in these proteins, cause development anomalies in eukaryotic organisms, from protozoa to mammals, indicating the physiological relevance of these transporters. In the SLC35A3 transporter, two mutations were associated with squeletal malformations and neurological characterized (Thomsen et al 2006; Edvardson 2013). In order to analyze the relation between SLC35A3 mutations and the congenital disorder of glycosylation (CDG), a pehotypic correction assay in yeast to functional characterization of the SLC35A3, WT and mutants, were designed. The study of mutants was based on bioinformatics tools, GFP-Fusion technology, a complementation activity assay in a yeast mutant and the use the lectine GSII-FITC to identified the GlcNAc residues. On the other hand the role of SLC35A3 on Xenopus laevis as a develompent model is been studied.The main advantages to work with Xenopus are, his common evolutionary history with mammals sharing extensive similarity at the level of the genomes and in organ development and the development outside of the progenitor of the embryos what makes them extremely well suited for manipulations of gene activity via microinjection (Christensen et al., 2008; Raciti et al., 2008). An expression profile using the In situ hybridization technique showed that the SLC35A3 from Xenopus laevis localize in ectodermal derivates cells. Further carry out knockdown studies by the use of morpholines will be clarifying to analyze the phenotypic results of the embryos.