In vivo trafficking and localization of p24 proteins in plant cells.
p24 proteins constitute a family of putative cargo receptors that traffic in the early secretory pathway. p24 proteins can be divided into four subfamilies (p23, p24, p25 and p26) by sequence homology. In contrast to mammals and yeast, most plant p24 proteins contain in their cytosolic C-terminus both a dilysine motif in the -3, -4 position and a diaromatic motif in the -7, -8 position. We have previously shown that the cytosolic tail of Arabidopsis p24 proteins has the ability to interact with ARF1 and coatomer (through the dilysine motif) and with COPII subunits (through the diaromatic motif). Here, we establish the localization and trafficking properties of an Arabidopsis thaliana p24 protein (Atp24) and have investigated the contribution of the sorting motifs in its cytosolic tail to its in vivo localization. Atp24-red fluorescent protein localizes exclusively to the endoplasmic reticulum (ER), in contrast with the localization of p24 proteins in other eukaryotes, and the dilysine motif is necessary and sufficient for ER localization. In contrast, Atp24 mutants lacking the dilysine motif are transported along the secretory pathway to the prevacuolar compartment and the vacuole, although a significant fraction is also found at the plasma membrane. Finally, we have found that ER export of Atp24 is COPII dependent, while its ER localization requires COPI function, presumably for efficient Golgi to ER recycling.
Langhans
Markus
M
Department of Cell Biology, Heidelberg Institute for Plant Sciences, University of Heidelberg, 69120 Heidelberg, Germany.
Marcote
María Jesús
MJ
Pimpl
Peter
P
Virgili-López
Goretti
G
Robinson
David G
DG
Aniento
Fernando
F
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D022181:COP-Coated Vesicles / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
C503841:ARF1 protein, Arabidopsis; D000199:Actins; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D020033:Protein Isoforms; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D008239:Lysine