Cloning and characterization of GNS1: a Saccharomyces cerevisiae gene involved in synthesis of 1,3-beta-glucan in vitro

The GNS1 gene product is required for the synthesis of 1,3-beta-glucan in vitro, since mutations in this gene result in exhibit an 80 to 90% reduction in 1,3-beta-glucan synthase specific activity. gns1 mutant strains display a pleiotropic phenotype including resistance to a pneumocandin B0 analog (...

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Bibliographic Details
Published in:Journal of Bacteriology 1995-06, Vol.177 (11), p.3227-3234
Main Authors: El-Sherbeini, M. (Merck Research Laboratories, Rahway, NJ.), Clemas, J.A
Format: Article
Language:English
Subjects:
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
Anti-Bacterial Agents - pharmacology
Bacteriology
Base Sequence
beta-Glucans
Chitin Synthase - metabolism
Cloning, Molecular
COMPOSICION QUIMICA
COMPOSITION CHIMIQUE
DNA Primers - chemistry
FENOTIPOS
Fungal Proteins - genetics
GENE
GENES
Genes, Fungal
Genetic Complementation Test
Genetics
GLICOSILTRANSFERASAS
GLUCANE
GLUCANO
Glucans - biosynthesis
Glucosyltransferases - genetics
GLYCOSYLTRANSFERASE
LOCI
LOCUS
Membrane Glycoproteins - genetics
METABOLISME DES GLUCIDES
METABOLISMO DE CARBOHIDRATOS
Molecular Sequence Data
MUTANT
MUTANTES
Mutation
Peptides
PHENOTYPE
Restriction Mapping
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - genetics
SECUENCIA NUCLEICA
SEQUENCE NUCLEIQUE
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Summary:The GNS1 gene product is required for the synthesis of 1,3-beta-glucan in vitro, since mutations in this gene result in exhibit an 80 to 90% reduction in 1,3-beta-glucan synthase specific activity. gns1 mutant strains display a pleiotropic phenotype including resistance to a pneumocandin B0 analog (L-733,560), slow growth, and mating and sporulation defects. The gns1-1 mutation was genetically mapped to within 135 centimorgans from the MAT locus on chromosome III. The wild-type GNS1 gene was isolated by complementing the pneomocandin resistance phenotype of the gns1-1 mutation and by hybridization with a chromosome III-derived sequence being used as a probe. The nucleotide sequence of GNS1 was determined and compared with the homologous region of the chromosome. The genetic and nucleotide sequence analyses revealed that GNS1 and the open reading frame, YCR34 [S. Oliver, Q. van der Aart, M. Agostoni-Carbone, and the Chromosome III Sequencing Group, Nature (London) 357:38-46, 1992], represent identical loci in the genome. Cells deleted for GNS1 are viable but exhibit slow growth as well as the pleiotropic phenotype of the gns1 mutants. The putative protein product is predicted to be an integral membrane protein with five transmembrane helices displaying an exoplasmic orientation for the N terminus and a cytoplasmic orientation for the C terminus. This protein may be a subunit of 1,3-beta-glucan synthase
ISSN:0021-9193
1098-5530
1067-8832
DOI:10.1128/jb.177.11.3227-3234.1995