Genomes and Genes
TRANSIENT GLUCOSYLATION OF GLYCOPROTEINS
Principal Investigator: ARMANDO JOSE PARODI
Abstract: DESCRIPTION (provided by applicant): The quality control (QC) of glycoprotein folding in the endoplasmic reticulum (ER) involves the interplay of a glucosyltransferase (GT) that only glycosylates not properly folded glycoprotein conformers, glucosidase II that removes residues added by GT and two ER resident lectins (calnexin, CNX and calreticulin, CRT) that specifically recognize monoglucosylated glycoproteins. This mechanism prevents exit of not properly folded glycoproteins to the Golgi and enhances glycoprotein folding efficiency and is directly related to the so-called "conformational diseases". To continue our characterization of the structural features that determine the participation of GT in the QC mechanism we now propose to study: (i) the subtle conformational differences, including those that may eventually result in the formation of amyloid fibers, that determine whether glycoproteins are recognized or not in vivo by GT;(ii) the maximum distance between the N-glycan and the structural distortion that allows GT-mediated glucosylation and (iii) whether glycoprotein structural stability determines its fate in the ER, i.e. successfully passing through the QC or being diverted to ER associated degradation (ERAD). For this purpose, we will express several N-glycosylation and stability lysozyme mutants (including some ones leading to the production of amyloids) in S. pombe cells and to follow whether they are recognized by GT, and whether they successfully fold or if, alternatively, they are derived to ERAD. These data, coupled to a structural characterization of the lysozyme mutants will provide information on why cells fail, in certain instances, to derive dangerous, aggregation-prone glycoproteins to degradation and on whether the structural stability of glycoproteins determines their fate in the ER. To continue our characterization of the role of N-glycans on ERAD we will further study the mechanism by which the ER mannosidase, the putative lectins Htm1p/Mnl1p/EDEM and Yos9p and the conformational sensor GT participate in driving irreparably misfolded glycoproteins to degradation. As a model system we chose S. pombe, a yeast that, contrary to what happens in S. cerevisiae, has a QC of glycoprotein folding similar to that occurring in mammalian cells. We plan to further study the proposal, derived from our own results, that the role of ER 1-mannosidase in ERAD is related to a putative lectin capacity of the protein and not to its enzymatic activity and to evaluate the relative importance in both productive glycoprotein folding and in diversion to ERAD of the first, partial glycan deglucosylation-dependent and of the following, reglucosylation, GT-dependent, CNX-glycoprotein interactions.
Funding Period: 1990-08-15 - 2012-07-31
more information: NIH RePORT
- UDP-GlC:glycoprotein glucosyltransferase-glucosidase II, the ying-yang of the ER quality controlCecilia D'Alessio
Laboratory of Glycobiology, Fundacion Instituto Leloir, Avda Patricias Argentinas 435, C1405BWE, Buenos Aires, Argentina
Semin Cell Dev Biol 21:491-9. 2010..This review deals with our present knowledge on the glucosyltransferase and the glucosidase...
- Functional cooperation between BiP and calreticulin in the folding maturation of a glycoprotein in Trypanosoma cruziCarlos A Labriola
Laboratories of Glycobiology, Fundación Instituto Leloir and Instituto de Investigaciones Bioquímicas de Buenos Aires IIBBA CONICET, Buenos Aires, Argentina
Mol Biochem Parasitol 175:112-7. 2011..The present report provides further evidence on the early evolutionary acquisition of the basic tenets of the N-glycan dependent quality control mechanism of glycoprotein folding...
- N-glycan trimming by glucosidase II is essential for Arabidopsis developmentPravina Soussilane
CNRS, UMR 6037, IFRMP 23, Bâtiment Biologie Extension, Faculte des Sciences, Mont Saint Aignan, France
Glycoconj J 26:597-607. 2009..Inactivation of the alpha subunit in a temperature sensitive Arabidopsis mutant blocked N-glycan processing after a first trimming by glucosidase I and strongly affected seedling development...
- How sugars convey information on protein conformation in the endoplasmic reticulumJulio J Caramelo
Laboratory of Glycobiology, Fundacion Instituto Leloir, Avda Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina
Semin Cell Dev Biol 18:732-42. 2007..The purpose of the review is to describe the most significant recent findings on the mechanism of glycoprotein folding and assembly quality control and to discuss the main still unanswered questions...