Isolation and Characterization of a Glycosylation Mutant from Schizosaccharomyces pombe

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 1996, Vol.60 (7), p.1156-1159
• Main Authors: Kaoru, Takegawa, Naotaka, Tanaka, Mitsuaki, Tabuchi, Shojiro, Iwahara
• Format: Article
• Language: English
• Subjects: Anti-Bacterial Agents - pharmacology; Biological and medical sciences; Biotechnology; Classical genetics, quantitative genetics, hybrids; Concanavalin A - chemistry; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; glactomannan; glycosylation; Glycosylation - drug effects; Hygromycin B - pharmacology; Lectins; Magnetic Resonance Spectroscopy; Mutation - physiology; Nlinked oligosaccharide; Oligosaccharides - genetics; Oligosaccharides - metabolism; Phenotype; Schizosaccharomyces - drug effects; Schizosaccharomyces - genetics; Schizosaccharomyces - metabolism; Schizosaccharomyces pombe; Thallophyta, bryophyta; Vanadates - pharmacology; Vegetals
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.60.1156

N-Linked oligosaccharides were elongated by glycosylation with mannose and galactose residues in the secretory pathway of Schizosaccharomyces pombe. The wild-type S. pombe cells were agglutinated by the additions of not only concanavalin A lectin, which is specific for mannose residues, but also PNA (from Arachis hypogaea) and RCA (Ricinus communis) lectins, which are specific for terminal galactose residues. By PNA-binding selection, we isolated an S. pombe mutant defective in protein glycosylation. The mutant cells, named gmsl, were not agglutinated by PNA or RCA. In contrast, agglutination of the gmsl cells by the addition of concanavalin A was markedly increased. Structural studies on N-linked oligosaccharides from gmsl mutant cells showed that the number of x-l,2-linked galactose residues wes markedly reduced, and unsubstituted x-l,6-linked polymannose outer chains were attached to the core oligosaccharides.