TY - JOUR
T1 - The fission yeast gmn2+ gene encodes an erd1 homologue of saccharomyces cerevisiae required for protein glycosylation and retention of luminal endoplasmic reticulum proteins
AU - Tanaka, Naotaka
AU - Kagami, Akinari
AU - Hirai, Keisuke
AU - Suzuki, Shotaro
AU - Matsuura, Shiori
AU - Fukunaga, Takamasa
AU - Tabuchi, Mitsuaki
AU - Takegawa, Kaoru
N1 - Funding Information:
We thank Drs. Clinton Ballou, Takashi Toda, Chikashi Shimoda, and John Armstrong for providing the S. pombe strains, antibodies, and plasmids. We acknowledge the technical expertise of the DNA core facility of the Gene Research Center, Kagawa University. This work was supported by JSPS KAKENHI Grant Numbers JP16K07669, and by the New Energy and Industrial Technology Development Organization (to K.T.).
Publisher Copyright:
© 2021 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation.
PY - 2021
Y1 - 2021
N2 - The gmn2 mutant of Schizosaccharomyces pombe has previously been shown to exhibit defects in protein glycosylation of N-linked oligosaccharides (Ballou, L. and Ballou, CE., Proc. Natl. Acad. Sci. USA, 92, 2790–2794 (1995)). Like most glycosylation-defective mutants, the S. pombe gmn2 mutant was found to be sensitive to hygromycin B, an aminoglycoside antibiotic. As a result of complementation analysis, the gmn2+ gene was found to be a single open reading frame that encodes a polypeptide of 373 amino acids consisting of multiple membrane-spanning regions. The Gmn2 protein shares sequence similarity with Kluyveromyces lactis and Saccharomyces cerevisiae Erd1 proteins, which are required for retention of luminal endoplasmic reticulum (ER) proteins. Although disruption of the gmn2+ gene is not lethal, the secreted glycoprotein showed a significant glycosylation defect with destabilization of the glycosyltransferase responsible for N-glycan elongation. It was also shown that a significant amount of BiP was missorted to the cell surface according to ADEL receptor destabilization. Fluorescent microscopy revealed that the functional Gmn2EGFP fusion protein is mainly localized in the Golgi membrane. These results indicate that the Gmn2 protein is required for protein glycosylation and for retention of ER-resident proteins in S. pombe cells.
AB - The gmn2 mutant of Schizosaccharomyces pombe has previously been shown to exhibit defects in protein glycosylation of N-linked oligosaccharides (Ballou, L. and Ballou, CE., Proc. Natl. Acad. Sci. USA, 92, 2790–2794 (1995)). Like most glycosylation-defective mutants, the S. pombe gmn2 mutant was found to be sensitive to hygromycin B, an aminoglycoside antibiotic. As a result of complementation analysis, the gmn2+ gene was found to be a single open reading frame that encodes a polypeptide of 373 amino acids consisting of multiple membrane-spanning regions. The Gmn2 protein shares sequence similarity with Kluyveromyces lactis and Saccharomyces cerevisiae Erd1 proteins, which are required for retention of luminal endoplasmic reticulum (ER) proteins. Although disruption of the gmn2+ gene is not lethal, the secreted glycoprotein showed a significant glycosylation defect with destabilization of the glycosyltransferase responsible for N-glycan elongation. It was also shown that a significant amount of BiP was missorted to the cell surface according to ADEL receptor destabilization. Fluorescent microscopy revealed that the functional Gmn2EGFP fusion protein is mainly localized in the Golgi membrane. These results indicate that the Gmn2 protein is required for protein glycosylation and for retention of ER-resident proteins in S. pombe cells.
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U2 - 10.2323/jgam.2020.07.002
DO - 10.2323/jgam.2020.07.002
M3 - Article
C2 - 33536395
AN - SCOPUS:85107711279
VL - 67
SP - 67
EP - 76
JO - Journal of General and Applied Microbiology
JF - Journal of General and Applied Microbiology
SN - 0022-1260
IS - 2
ER -