Overexpression of Arabidopsis sorting nexin AtSNX2b inhibits endocytic trafficking to the vacuole.
Sorting nexins are conserved proteins that function in vesicular trafficking and contain a characteristic phox homology (PX) domain. Here, we characterize the ubiquitously expressed Arabidopsis thaliana sorting nexin AtSNX2b. Sub-cellular fractionation studies indicate that AtSNX2b is peripherally associated with membranes. The AtSNX2b PX domain binds to phosphatidylinositol 3-phosphate in vitro and this association is required for the localization of GFP-AtSNX2b to punctate structures in vivo, identified as the trans-Golgi network, prevacuolar compartment and endosomes. Overexpression of GFP-tagged AtSNX2b produces enlarged GFP-labeled compartments that can also be labeled by the endocytic tracer FM4-64. Endocytic trafficking of FM4-64 to the vacuole is arrested in these GFP-AtSNX2b compartments, and similar FM4-64-accumulating compartments are seen upon overexpression of untagged AtSNX2b. This suggests that exit of membrane components from these enlarged or aggregated endosomes is inhibited. Vacuolar proteins containing an N-terminal propeptide, but not those with a C-terminal propeptide, are also present in these enlarged compartments. We hypothesize that AtSNX2b is involved in vesicular trafficking from endosomes to the vacuole.
Phan
Nguyen Q
NQ
Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA.
Kim
Sang-Jin
SJ
Bassham
Diane C
DC
D017360:Arabidopsis / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D004705:Endocytosis / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D011523:Protoplasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D058305:Sorting Nexins; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014617:Vacuoles / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000737:chemistry / Q000378:metabolism*
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D018129:Phosphatidylinositol Phosphates; D011993:Recombinant Fusion Proteins; C546322:SNX2b protein, Arabidopsis; D058305:Sorting Nexins; D033921:Vesicular Transport Proteins; C055525:phosphatidylinositol 3-phosphate; D049452:Green Fluorescent Proteins