Novel type Arabidopsis thaliana H(+)-PPase is localized to the Golgi apparatus.
Vacuolar H(+)-PPase, a membrane bound proton-translocating pyrophosphatase found in various species including plants, some protozoan and prokaryotes, has been demonstrated to be localized to the vacuolar membrane in plants. Using a GUS reporter system and a green fluorescent protein (GFP) fusion protein, we investigated the tissue distribution and the subcellular localization, respectively, of a novel type H(+)-PPase encoded by AVP2/AVPL1 identified in the Arabidopsis thaliana genome. We showed that AVP2/AVPL1 is highly expressed at the trichome and the filament of stamen. Furthermore, the fluorescence of GFP-tagged AVP2/AVPL1 showed small dot-like structures that were observed throughout the cytoplasm of various Arabidopsis cells under a fluorescent microscope. The distribution of this dot-like fluorescent pattern was apparently affected by a treatment with brefeldin A. Moreover, we demonstrated that most dot-like fluorescent structures colocalized with a Golgi resident protein. These findings suggest that this novel type H(+)-PPase resides on the Golgi apparatus rather than the vacuolar membrane.
Mitsuda
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N
Graduate School of Human and Environmental Studies, Kyoto University, Kyoto, Japan.
Enami
K
K
Nakata
M
M
Takeyasu
K
K
Sato
M H
MH
D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D020126:Brefeldin A / Q000494:pharmacology; D003593:Cytoplasm / Q000187:drug effects / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter / Q000235:genetics; D006056:Golgi Apparatus / Q000187:drug effects / Q000201:enzymology*; D043564:Inorganic Pyrophosphatase; D007425:Intracellular Membranes / Q000201:enzymology; D008856:Microscopy, Fluorescence; D018514:Plant Structures / Q000201:enzymology; D011401:Promoter Regions, Genetic / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000201:enzymology
D011993:Recombinant Fusion Proteins; D020126:Brefeldin A; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase