The plant vesicle-associated SNARE AtVTI1a likely mediates vesicle transport from the trans-Golgi network to the prevacuolar compartment.
Membrane traffic in eukaryotic cells relies on recognition between v-SNAREs on transport vesicles and t-SNAREs on target membranes. Here we report the identification of AtVTI1a and AtVTI1b, two Arabidopsis homologues of the yeast v-SNARE Vti1p, which is required for multiple transport steps in yeast. AtVTI1a and AtVTI1b share 60% amino acid identity with one another and are 32 and 30% identical to the yeast protein, respectively. By suppressing defects found in specific strains of yeast vti1 temperature-sensitive mutants, we show that AtVTI1a can substitute for Vti1p in Golgi-to-prevacuolar compartment (PVC) transport, whereas AtVTI1b substitutes in two alternative pathways: the vacuolar import of alkaline phosphatase and the so-called cytosol-to-vacuole pathway used by aminopeptidase I. Both AtVTI1a and AtVTI1b are expressed in all major organs of Arabidopsis. Using subcellular fractionation and immunoelectron microscopy, we show that AtVTI1a colocalizes with the putative vacuolar cargo receptor AtELP on the trans-Golgi network and the PVC. AtVTI1a also colocalizes with the t-SNARE AtPEP12p to the PVC. In addition, AtVTI1a and AtPEP12p can be coimmunoprecipitated from plant cell extracts. We propose that AtVTI1a functions as a v-SNARE responsible for targeting AtELP-containing vesicles from the trans-Golgi network to the PVC, and that AtVTI1b is involved in a different membrane transport process.
Department of Energy-Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48824, USA.
H
H
Zheng
G F
GF
von Mollard
V
V
Kovaleva
T H
TH
Stevens
N V
NV
Raikhel
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D002451:Cell Compartmentation; D000939:Epitopes; D018506:Gene Expression Regulation, Plant; D006056:Golgi Apparatus / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011233:Precipitin Tests; D050766:Qb-SNARE Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013395:Sucrose; D014158:Transcription, Genetic; D014461:Ultracentrifugation; D014617:Vacuoles / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D029681:Arabidopsis Proteins; D002352:Carrier Proteins; C106564:ELP protein, Arabidopsis; D000939:Epitopes; D008565:Membrane Proteins; D010940:Plant Proteins; D050766:Qb-SNARE Proteins; D011994:Recombinant Proteins; C120751:vesicle transport v-SNARE 11 protein, Arabidopsis; C120752:vesicle transport v-SNARE 12 protein, Arabidopsis; D013395:Sucrose