title: Isolation of Sporothrix schenckii GDA1 and functional characterization of the encoded guanosine diphosphatase activity.
abstract: Sporothrix schenckii is a fungal pathogen of humans and the etiological agent of sporotrichosis. In fungi, proper protein glycosylation is usually required for normal composition of cell wall and virulence. Upon addition of precursor oligosaccharides to nascent proteins in the endoplasmic reticulum, glycans are further modified by Golgi-glycosyl transferases. In order to add sugar residues to precursor glycans, nucleotide diphosphate sugars are imported from the cytosol to the Golgi lumen, the sugar is transferred to glycans, and the resulting nucleoside diphosphate is dephosphorylated by the nucleoside diphosphatase Gda1 before returning to cytosol. Here, we isolated the open reading frame SsGDA1 from a S. schenckii genomic DNA library. In order to confirm the function of SsGda1, we performed complementation assays in a Saccharomyces cerevisiae gda1∆ null mutant. Our results indicated that SsGDA1 restored the nucleotide diphosphatase activity to wild-type levels and therefore is a functional ortholog of S. cerevisiae GDA1.
authors.affiliation: Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, Col. Noria Alta, CP 36050, Guanajuato, Gto, Mexico.
authors.forename: Adolfo
authors.initials: A
authors.lastname: López-Esparza
authors.forename: Aurelio
authors.initials: A
authors.lastname: Álvarez-Vargas
authors.forename: Héctor M
authors.initials: HM
authors.lastname: Mora-Montes
authors.forename: Arturo
authors.initials: A
authors.lastname: Hernández-Cervantes
authors.forename: Ma
authors.initials: M
authors.lastname: Del Carmen Cano-Canchola
authors.forename: Arturo
authors.initials: A
authors.lastname: Flores-Carreón
mesh_terms: D000595:Amino Acid Sequence; D002473:Cell Wall / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D008969:Molecular Sequence Data; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013175:Sporothrix / Q000201:enzymology* / Q000235:genetics*
chemical_list: D005656:Fungal Proteins; D011755:Pyrophosphatases; C010357:guanosine-diphosphatase