title: Identification of alg3 in the mushroom-forming fungus Schizophyllum commune and analysis of the Δalg3 knockout mutant.
abstract: Alg3 of Saccharomyces cerevisiae catalyzes the mannosyl transfer from Man-P-Dol to Man(5)GlcNAc(2)-PP-Dol resulting in the formation of Man(6)GlcNAc(2)-PP-Dol, which is then further processed to the final precursor oligosaccharide Glc(3)Man(9)GlcNAc(2) for N-glycosylation of proteins. Here, we identified the alg3 gene of the mushroom-forming fungus Schizophyllum commune by homology search. Its function was confirmed by the complementation of the Δalg3 strain of S. cerevisiae. Inactivation of alg3 in S. commune resulted in the production of predominantly Man(3)GlcNAc(2) protein-linked N-glycans. No impact on growth nor a developmental phenotype due to the deletion was observed. This provides a first step toward engineering of a homogeneous, humanized N-glycosylation pattern for the production of therapeutic glycoproteins in mushrooms.
authors.affiliation: Microbiology and Kluyver Centre for Genomics of Industrial Fermentations, Utrecht University, Utrecht, The Netherlands. e.berends@uu.nl
authors.forename: Elsa
authors.initials: E
authors.lastname: Berends
authors.forename: Ludwig
authors.initials: L
authors.lastname: Lehle
authors.forename: Maurice
authors.initials: M
authors.lastname: Henquet
authors.forename: Thamara
authors.initials: T
authors.lastname: Hesselink
authors.forename: Han A B
authors.initials: HA
authors.lastname: Wösten
authors.forename: Luis G
authors.initials: LG
authors.lastname: Lugones
authors.forename: Dirk
authors.initials: D
authors.lastname: Bosch
mesh_terms: D000363:Agaricales / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D000595:Amino Acid Sequence; D055786:Gene Knockout Techniques; D006023:Glycoproteins / Q000096:biosynthesis*; D006031:Glycosylation; D008364:Mannosyltransferases / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D012567:Schizophyllum / Q000235:genetics / Q000378:metabolism
chemical_list: D006023:Glycoproteins; C503888:Man(3)GlcNAc(2); D008565:Membrane Proteins; D009844:Oligosaccharides; D029701:Saccharomyces cerevisiae Proteins; C104466:ALG3 protein, S cerevisiae; D008364:Mannosyltransferases