SpMnn9p and SpAnp1p form a protein complex involved in mannan synthesis in the fission yeast Schizosaccharomyces pombe
Section snippets
Strains, media and genetic methods
Wild-type S. pombe ARC039 (h-ura4-C190T leu1-32) strain was provided by the National BioResource Project (NBRP). Standard rich medium (YES) and synthetic minimal medium (MM) were used for growing S. pombe as described (29). For transformation of S. pombe cells, the lithium acetate method was used (30,31). Standard methods used for genetic manipulation have been described previously (32). Escherichia coli XL1-Blue (Stratagene, La Jolla, CA, USA) was used for all cloning procedures.
Gene disruption
The Spmnn9+
Phenotypic characterization of Spmnn9Δ and Spanp1Δ cells
To investigate the effects of deletions of Spmnn9+ and Spanp1+ genes on the cell growth and glycosylation, Spmnn9Δ and Spanp1Δ cells were constructed. Both Spmnn9Δ and Spanp1Δ cells were aberrantly shaped and swollen, and appeared to be inherently aggregating in cultures with approximately 5–10 cells, similar to the Spoch1Δ cells (39) (Fig. 1A). EDTA treatment, which is used for disaggregating flocculation mutants (40), did not abolish the aggregations of the Spmnn9Δ and the Spanp1Δ cells (data
Discussion
The mechanism underlying the biosynthesis of mannan structure in S. cerevisiae has been extensively investigated, leading to the identification of the genes involved in mannan biosynthesis and elucidation of the coordinated structure of mannan. However, molecular details of the mechanism of mannan biosynthesis in S. pombe still remains largely elusive, even though the genes homologous to those involved in mannan biosynthesis in S. cerevisiae were predicted to be present in S. pombe. Here we
Acknowledgments
This work was partly supported by the Project for Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers by the Ministry of Economy, Trade and Industry (METI) of Japan, New Energy and Industrial Technology Development Organization (NEDO).
References (51)
- et al.
Overview of N- and O-linked oligosaccharide structures found in various yeast species
Biochim. Biophys. Acta
(1999) - et al.
The fission yeast Pvg1p has galactose-specific pyruvytransferase activity
FEBS Lett.
(2013) - et al.
Characterization of N- and O-linked galactosylated oligosaccharides from fission yeast species
J. Biosci. Bioeng.
(2020) - et al.
Schizosaccharomyces pombe och1+ encodes α-1,6-mannosyltransferase that is involved in outer chain elongation of N-linked oligosaccharides
FEBS Lett.
(2001) - et al.
Novel membrane protein complexes for protein glycosylation in the yeast Golgi apparatus
Biochem. Biophys. Res. Commun.
(1997) - et al.
Expression and purification of recombinant M-Pol I from Saccharomyces cerevisiae with α1,6-mannosyltransferase activity
Protein Expr. Purif.
(2009) - et al.
The components of the Saccharomyces cerevisiae mannosyltransferase complex M-Pol I have distinct functions in mannan synthesis
J. Biol. Chem.
(2002) - et al.
Identification of the MNN2 and MNN5 mannosyltransferases required for forming and extending the mannose branches of the outer chain mannans of Saccharomyces cerevisiae
J. Biol. Chem.
(1998) - et al.
The KTR and MNN1 mannosyltransferase families of Saccharomyces cerevisiae
Biochim. Biophys. Acta
(1999) - et al.
Mannosylphosphate transfer to yeast mannan
Biochim. Biophys. Acta
(1999)
Molecular genetic analysis of fission yeast Schizosaccharomyces pombe
Methods Enzymol.
Structure analysis of oligosaccharides by tagging of the reducing end sugars with a fluorescent compound
Biochem. Biophys. Res. Commun.
High-performance liquid chromatography of pyridylaminated saccharides
Methods Enzymol.
The evolution of two-dimensional gel electrophoresis – from proteomics to emerging alternative applications
J. Chromatogr. A
Production of heterologous glycoproteins by a glycosylation-defective alg3och1 mutant of Schizosaccharomyces pombe
J. Biotechnol.
OCH1 encodes a novel membrane bound mannosyltransferase: outer chain elongation of asparagine-linked oligosaccharides
EMBO J.
Multi-protein complexes in the cis Golgi of Saccharomyces cerevisiae with α-1,6-mannosyltransferase activity
EMBO J.
Yeast Mnn9 is both a priming glycosyltransferase and an allosteric activator of mannan biosynthesis
Open Biol.
Interaction among the subunits of Golgi membrane mannosyltransferase complexes of the yeast Saccharomyces cerevisiae
Biosci. Biotechnol. Biochem.
The Saccharomyces cerevisiae protein Mnn10p/Bed1p is a subunit of a Golgi mannosyltransferase complex
J. Biol. Chem.
Active recycling of yeast Golgi mannosyltransferase complexes though the endoplasmic reticulum
Proc. Natl. Acad. Sci. USA
The cytoplasmic region of α-1,6-mannosyltransferase Mnn9p is crucial for retrograde transport from the Golgi apparatus to the endoplasmic reticulum in Saccharomyces cerevisiae
Eukaryot. Cell
The functioning of the yeast Golgi apparatus requires an ER protein encoded by ANP1, a member of a new family of genes affecting the secretory pathway
EMBO J.
Cloning and analysis of the S. cerevisiae MNN9 and MNN1 genes required for complex glycosylation of secreted proteins
Proc. Natl. Acad. Sci. USA
Sodium orthovanadate-resistant mutants of Saccharomyces cerevisiae show defects in Golgi-mediated protein glycosylation, sporulation and detergent resistance
Genetics
Cited by (6)
Yeast engineering technologies and their applications to the food industry
2021, Food BiotechnologyThe fission yeast gmn2<sup>+</sup> gene encodes an erd1 homologue of saccharomyces cerevisiae required for protein glycosylation and retention of luminal endoplasmic reticulum proteins
2021, Journal of General and Applied MicrobiologyGolgi localization of glycosyltransferases requires Gpp74p in Schizosaccharomyces pombe
2020, Applied Microbiology and Biotechnology
- §
Present address: Department of Life Science, Faculty of Science and Engineering, Setsunan University, 17-8 Ikeda-Nakamachi, Neyagawa, Osaka 572–8508, Japan.