A vacuolar carboxypeptidase mutant of Arabidopsis thaliana is degraded by the ERAD pathway independently of its N-glycan

Masaya Yamamoto, Mitsuyoshi Kawanabe, Yoko Hayashi, Toshiya Endo, Shuh ichi Nishikawa

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Misfolded proteins produced in the endoplasmic reticulum (ER) are degraded by a mechanism, the ER-associated degradation (ERAD). Here we report establishment of the experimental system to analyze the ERAD in plant cells. Carboxypeptidase Y (CPY) is a vacuolar enzyme and its mutant CPY* is degraded by the ERAD in yeast. Since Arabidopsis thaliana has AtCPY, an ortholog of yeast CPY, we constructed and expressed fusion proteins consisting of AtCPY and GFP and of AtCPY*, which carries a mutation homologous to yeast CPY*, and GFP in A. thaliana cells. While AtCPY-GFP was efficiently transported to the vacuole, AtCPY*-GFP was retained in the ER to be degraded in proteasome- and Cdc48-dependent manners. We also found that AtCPY*-GFP was degraded by the ERAD in yeast cells, but that its single N-glycan did not function as a degradation signal in yeast or plant cells. Therefore, AtCPY*-GFP can be used as a marker protein to analyze the ERAD pathway, likely for nonglycosylated substrates, in plant cells.

Original languageEnglish
Pages (from-to)384-389
Number of pages6
JournalBiochemical and biophysical research communications
Volume393
Issue number3
DOIs
Publication statusPublished - 2010 Mar 12
Externally publishedYes

Keywords

  • Arabidopsis thaliana
  • ER-associated degradation (ERAD)
  • Endoplasmic reticulum (ER)
  • Serine carboxypeptidase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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