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N-glycans are not required for the efficient degradation of the mutant Saccharomyces cerevisiae CPY* in Schizosaccharomyces pombe

  • Applied genetics and molecular biotechnology
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Abstract

In eukaryotic cells, aberrant proteins generated in the endoplasmic reticulum (ER) are degraded by the ER-associated degradation (ERAD) pathway. Here, we report on the ERAD pathway of the fission yeast Schizosaccharomyces pombe. We constructed and expressed Saccharomyces cerevisiae wild-type CPY (ScCPY) and CPY-G255R mutant (ScCPY*) in S. pombe. While ScCPY was glycosylated and efficiently transported to the vacuoles in S. pombe, ScCPY* was retained in the ER and was not processed to the matured form in these cells. Cycloheximide chase experiments revealed that ScCPY* was rapidly degraded in S. pombe, and its degradation depended on Hrd1p and Ubc7p homologs. We also found that Mnl1p and Yos9p, proteins that are essential for ERAD in S. cerevisiae, were not required for ScCPY* degradation in S. pombe. Moreover, the null-glycosylation mutant of ScCPY, CPY*0000, was rapidly degraded by the ERAD pathway. These results suggested that N-linked oligosaccharides are not important for the recognition of luminal proteins for ERAD in S. pombe cells.

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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 & Industry, as supported by the New Energy and Industrial Technology Development Organization.

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Authors and Affiliations

  1. ASPEX Division, Research Center, Asahi Glass Co., Ltd, 1150 Hazawa-cho, Kanagawa, 221-8755, Japan

    Hiroyuki Mukaiyama

  2. Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan

    Hiroyuki Mukaiyama, Michiko Kodera, Naotaka Tanaka & Kaoru Takegawa

  3. Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Fukuoka, 812-8581, Japan

    Hiroyuki Mukaiyama & Kaoru Takegawa

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  1. Hiroyuki Mukaiyama
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  2. Michiko Kodera
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  3. Naotaka Tanaka
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  4. Kaoru Takegawa
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Correspondence to Kaoru Takegawa.

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Mukaiyama, H., Kodera, M., Tanaka, N. et al. N-glycans are not required for the efficient degradation of the mutant Saccharomyces cerevisiae CPY* in Schizosaccharomyces pombe . Appl Microbiol Biotechnol 93, 1609–1618 (2012). https://doi.org/10.1007/s00253-011-3662-z

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  • DOI: https://doi.org/10.1007/s00253-011-3662-z

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