A vacuolar carboxypeptidase mutant of Arabidopsis thaliana is degraded by the ERAD pathway independently of its N-glycan
In: Biochemical and Biophysical Research Communications, Jg. 393 (2010-03-01), S. 384-389
Online
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Zugriff:
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.
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A vacuolar carboxypeptidase mutant of Arabidopsis thaliana is degraded by the ERAD pathway independently of its N-glycan
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Autor/in / Beteiligte Person: | Kawanabe, Mitsuyoshi ; Endo, Toshiya ; Yamamoto, Masaya ; Nishikawa, Shuh-ichi ; Hayashi, Yoko |
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Zeitschrift: | Biochemical and Biophysical Research Communications, Jg. 393 (2010-03-01), S. 384-389 |
Veröffentlichung: | Elsevier BV, 2010 |
Medientyp: | unknown |
ISSN: | 0006-291X (print) |
DOI: | 10.1016/j.bbrc.2010.02.001 |
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