title: Cloning and characterization of cDNA of the GPI-anchored purple acid phosphatase and its root tissue distribution in Spirodela oligorrhiza.
abstract: A cDNA clone of the glycosylphosphatidylinositol (GPI)-anchored purple acid phosphatase (PAP) has been obtained by a combination of cDNA library screening and 5' rapid amplification of cDNA ends from Spirodela oligorrhiza plants grown under phosphate-deficient (-P) conditions. The open reading frame of the S. oligorrhiza PAP cDNA consists of 1 365 bp encoding a 455 amino acid protein. Its deduced amino acid sequence shows 82 and 80% similarity to Arabidopsis thaliana and Phaseolus vulgaris PAP, respectively. The amino acid residue, Ala439, followed by two more small amino acid residues, Asp and Ser, is predicted to be the GPI-anchoring (omega) site. The absence of a dibasic motif upstream of the putative omega site suggests that the PAP is a cell wall protein. This presumption is supported by the finding that PAP was released by digestion of the cell wall fraction with cellulase. The GPI anchor is speculated to be a signal for transporting PAP to the cell wall. Immunohistochemical results using -P plant roots demonstrate that PAP is preferentially distributed in the outermost cortical cells of roots but not in the epidermis, suggesting its role in acquiring inorganic phosphate under phosphate-deficient conditions. Northern blot analysis using the S. oligorrhiza PAP cDNA as a probe demonstrates that expression of the PAP gene increased during growth of -P plants and this time-dependent occurrence in mRNA levels of the PAP in -P plants was also observed in their protein and activity levels.
authors.affiliation: Laboratory of Environmental Molecular Biology, Graduate School of Environmental Earth Science, Hokkaido University, Kita-ku, Sapporo 060-0810, Japan Biological Laboratory, Hakodate College, Hokkaido University of Education, Hachiman-cho, Hakodate 040-8567, Japan Molecular Adaptation Research Group, Research Institute of Biological Resources, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-higashi, Toyohira-ku, Sapporo 062-8517, Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Nishikoori
authors.forename: Kenji
authors.initials: K
authors.lastname: Washio
authors.forename: Akira
authors.initials: A
authors.lastname: Hase
authors.forename: Naoki
authors.initials: N
authors.lastname: Morita
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Okuyama