title: Functional characterization of Sporothrix schenckii glycosidases involved in the N-linked glycosylation pathway.
abstract: Protein glycosylation pathways are conserved metabolic processes in eukaryotic organisms and are required for cell fitness. In fungal pathogens, the N-linked glycosylation pathway is indispensable for proper cell wall composition and virulence. In Sporothrix schenckii sensu stricto, the causative agent of sporotrichosis, little is known about this glycosylation pathway. Here, using a genome-wide screening for putative members of the glycosyl hydrolase (CAZy - GH) families 47 and 63, which group enzymes involved in the processing step during N-linked glycan maturation, we found seven homologue genes belonging to family 47 and one to family 63. The eight genes were individually expressed in C. albicans null mutants lacking either MNS1 (for members of family 47) or CWH41 (for the member of family 63). Our results indicate that SsCWH41 is the functional ortholog of CaCWH41, whereas SsMNS1 is the functional ortholog of CaMNS1. The remaining genes of family 47 encode Golgi mannosidases and endoplasmic reticulum degradation-enhancing alpha-mannosidase-like proteins (EDEMs). Since these GH families gather proteins used as target for drugs to control cell growth, identification of these genes could help in the design of antifungals that could be used to treat sporotrichosis and other fungal diseases. In addition, to our knowledge, we are the first to report that Golgi mannosidases and EDEMs are expressed and characterized in yeast cells.
authors.affiliation: Departamento de Biologia Celular, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Leila M
authors.initials: LM
authors.lastname: Lopes-Bezerra
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, colonia Noria Alta, Guanajuato, Guanajuato, México.
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Lozoya-Pérez
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, colonia Noria Alta, Guanajuato, Guanajuato, México.
authors.forename: Luz A
authors.initials: LA
authors.lastname: López-Ramírez
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, colonia Noria Alta, Guanajuato, Guanajuato, México.
authors.forename: José A
authors.initials: JA
authors.lastname: Martínez-Álvarez
authors.affiliation: Departamento de Biologia Celular, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
authors.forename: Marcus M
authors.initials: MM
authors.lastname: Teixeira
authors.affiliation: Departamento de Biologia Celular, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
authors.forename: Maria S S
authors.initials: MS
authors.lastname: Felipe
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, colonia Noria Alta, Guanajuato, Guanajuato, México.
authors.forename: Arturo
authors.initials: A
authors.lastname: Flores-Carreón
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, colonia Noria Alta, Guanajuato, Guanajuato, México hmora@ugto.mx.
authors.forename: Héctor M
authors.initials: HM
authors.lastname: Mora-Montes
mesh_terms: D002176:Candida albicans / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D005656:Fungal Proteins / Q000378:metabolism*; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D013175:Sporothrix / Q000201:enzymology* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D006026:Glycoside Hydrolases
keywords: Candida albicans; N-linked glycosylation; Sporothrix schenckii; glycosidases; heterologous expression