Novel type Arabidopsis thaliana H +-PPase is localized to the Golgi apparatus

• Published in: FEBS letters 2001-01, Vol.488 (1), p.29-33
• Main Authors: Mitsuda, Nobutaka, Enami, Kazuhiko, Nakata, Mami, Takeyasu, Kunio, Sato, Masa H.
• Format: Article
• Language: English
• Subjects: Arabidopsis - cytology; Arabidopsis - drug effects; Arabidopsis - enzymology; Arabidopsis thaliana; AVP2/AVPL1 gene; Brefeldin A; Brefeldin A - pharmacology; Cytoplasm - drug effects; Cytoplasm - enzymology; Fluorescent Antibody Technique; Gene Expression Regulation, Plant; Genes, Reporter - genetics; GFP, green fluorescent protein; Golgi apparatus; Golgi Apparatus - drug effects; Golgi Apparatus - enzymology; Green fluorescent protein; H +-PPase; Inorganic Pyrophosphatase; Intracellular Membranes - enzymology; Microscopy, Fluorescence; MS, Murashige–Skoog; Plant Structures - enzymology; Promoter Regions, Genetic - genetics; Protein Transport - drug effects; Pyrophosphatases - chemistry; Pyrophosphatases - genetics; Pyrophosphatases - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism; Vacuoles - drug effects; Vacuoles - enzymology
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(00)02400-5

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.