Role of the leader sequence in tobacco pectin methylesterase secretion.
We report that unprocessed tobacco pectin methylesterase (PME) contains N-terminal pro-sequence including the transmembrane (TM) domain and spacer segment preceding the mature PME. The mature portion of PME was replaced by green fluorescent protein (GFP) gene and various deletion mutants of pro-sequence fused to GFP were cloned into binary vectors and agroinjected in Nicotiana benthamiana leaves. The PME pro-sequence delivered GFP to the cell wall (CW). We showed that a transient binding of PME TM domain to endoplasmic reticulum membranes occurs upon its transport to CW. The CW targeting was abolished by various deletions in the TM domain, i.e., anchor domain was essential for secretion of GFP to CW. By contrast, even entire deletion of the spacer segment had no influence on GFP targeting.
Dorokhov
Yuri L
YL
Department of Virology, A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Leninsky Gory 1, Laboratory Building A, Moscow 119992, Russia.
Skurat
Eugene V
EV
Frolova
Olga Yu
OY
Gasanova
Tatjana V
TV
Ivanov
Peter A
PA
Ravin
Nikolay V
NV
Skryabin
Konstantin G
KG
Mäkinen
Kristiina M
KM
Klimyuk
Viktor I
VI
Gleba
Yuri Yu
YY
Atabekov
Joseph G
JG
D000595:Amino Acid Sequence; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D017343:Genes, Plant; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology*; D017384:Sequence Deletion; D014026:Nicotiana / Q000201:enzymology*
D010940:Plant Proteins; D021382:Protein Sorting Signals; D049452:Green Fluorescent Proteins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase