Protein recycling from the Golgi apparatus to the endoplasmic reticulum in plants and its minor contribution to calreticulin retention.
Using pulse-chase experiments combined with immunoprecipitation and N-glycan structural analysis, we showed that the retrieval mechanism of proteins from post-endoplasmic reticulum (post-ER) compartments is active in plant cells at levels similar to those described previously for animal cells. For instance, recycling from the Golgi apparatus back to the ER is sufficient to block the secretion of as much as 90% of an extracellular protein such as the cell wall invertase fused with an HDEL C-terminal tetrapeptide. Likewise, recycling can sustain fast retrograde transport of Golgi enzymes into the ER in the presence of brefeldin A. However, on the basis of our data, we propose that this retrieval mechanism in plants has little impact on the ER retention of a soluble ER protein such as calreticulin. Indeed, the latter is retained in the ER without any N-glycan-related evidence for a recycling through the Golgi apparatus. Taken together, these results indicate that calreticulin and perhaps other plant reticuloplasmins are possibly largely excluded from vesicles exported from the ER. Instead, they are probably retained in the ER by mechanisms that rely primarily on signals other than H/KDEL motifs.
Pagny
S
S
Laboratoire des Transports Intracellulaires, CNRS-ESA 6037, IFRMP23, Université de Rouen, 76821 Mont St. Aignan Cedex, France.
Cabanes-Macheteau
M
M
Gillikin
J W
JW
Leborgne-Castel
N
N
Lerouge
P
P
Boston
R S
RS
Faye
L
L
Gomord
V
V
D001483:Base Sequence; D020126:Brefeldin A / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D037282:Calreticulin; D017931:DNA Primers; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D006868:Hydrolysis; D008358:Mannose / Q000032:analysis; D010940:Plant Proteins / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis; D011499:Protein Processing, Post-Translational; D012261:Ribonucleoproteins / Q000737:chemistry / Q000378:metabolism*; D003313:Zea mays / Q000737:chemistry; D043324:beta-Fructofuranosidase
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D017931:DNA Primers; D010940:Plant Proteins; D011134:Polysaccharides; D012261:Ribonucleoproteins; D020126:Brefeldin A; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D008358:Mannose