Ricin A chain without its partner B chain is degraded after retrotranslocation from the endoplasmic reticulum to the cytosol in plant cells.
When expressed in tobacco cells, the catalytic subunit of the dimeric ribosome inactivating protein, ricin, is first inserted into the endoplasmic reticulum (ER) and then degraded in a manner that can be partially inhibited by the proteasome inhibitor clasto-lactacystin beta-lactone. Consistent with the implication of cytosolic proteasomes, degradation of ricin A chain is brefeldin A-insensitive and the polypeptides that accumulate in the presence of the proteasome inhibitor are not processed in a vacuole-specific fashion. Rather, these stabilized polypeptides are in part deglycosylated by a peptide:N-glycanase-like activity. Taken together, these results indicate that ricin A chain, albeit a structurally native protein, can behave as a substrate for ER to cytosol export, deglycosylation in the cytosol, and proteasomal degradation. Furthermore, retrotranslocation of this protein is not tightly coupled to proteasomal activity. These data are consistent with the hypothesis that ricin A chain can exploit the ER-associated protein degradation pathway to reach the cytosol. Although well characterized in mammalian and yeast cells, the operation of a similar pathway to the cytosol of plant cells has not previously been demonstrated.
Di Cola
A
A
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom.
Frigerio
L
L
Lord
J M
JM
Ceriotti
A
A
Roberts
L M
LM
D001693:Biological Transport, Active; D020126:Brefeldin A / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007783:Lactones / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism; D037121:Plant Lectins; D030821:Plants, Genetically Modified; D046988:Proteasome Endopeptidase Complex; D021122:Protein Subunits; D012276:Ricin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000378:metabolism*
D015853:Cysteine Proteinase Inhibitors; D007783:Lactones; D009097:Multienzyme Complexes; D037121:Plant Lectins; D021122:Protein Subunits; C098869:clasto-lactacystin beta-lactone; D020126:Brefeldin A; D012276:Ricin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex