title: Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum.
abstract: Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum.
authors.affiliation: Departamento de Genetica, Universidad de Cordoba, Spain; Campus de Excelencia Agroalimentario (ceiA3), E-14071 Cordoba, Spain.
authors.forename: Loida
authors.initials: L
authors.lastname: Lopez-Fernandez
authors.affiliation: Departamento de Genetica, Universidad de Cordoba, Spain; Campus de Excelencia Agroalimentario (ceiA3), E-14071 Cordoba, Spain.
authors.forename: M Isabel G
authors.initials: MI
authors.lastname: Roncero
authors.affiliation: Centro de Investigaciones Biologicas-CSIC, 28040 Madrid, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Prieto
authors.affiliation: Departamento de Genetica, Universidad de Cordoba, Spain; Campus de Excelencia Agroalimentario (ceiA3), E-14071 Cordoba, Spain. Electronic address: ge2rurom@uco.es.
authors.forename: Carmen
authors.initials: C
authors.lastname: Ruiz-Roldan
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D005670:Fusarium / Q000378:metabolism*; D020869:Gene Expression Profiling; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods; D020543:Proteome / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D008565:Membrane Proteins; D011134:Polysaccharides; D020543:Proteome; D016695:Glycosyltransferases
keywords: Cell wall structure; Fusarium; N-glycosylation; Protein secretion; Virulence