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Role of the leader sequence in tobacco pectin methylesterase secretion
Author(s) -
Dorokhov Yuri L.,
Skurat Eugene V.,
Frolova Olga Yu.,
Gasanova Tatjana V.,
Ivanov Peter A.,
Ravin Nikolay V.,
Skryabin Konstantin G.,
Mäkinen Kristiina M.,
Klimyuk Viktor I.,
Gleba Yuri Yu.,
Atabekov Joseph G.
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.04.090
Subject(s) - green fluorescent protein , nicotiana benthamiana , endoplasmic reticulum , secretion , transmembrane domain , chemistry , microbiology and biotechnology , mutant , biochemistry , biophysics , biology , gene
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.