the arabidopsis apyrase atapy1 is localized in the golgi instead of the extracellular space的拟南芥apyrase atapy1在高尔基体而不是局部细胞外空间.pdfVIP

The Arabidopsis apyrase AtAPY1 is localized in the Golgi instead of the extracellular space Schiller et al. Schiller et al. BMC Plant Biology 2012, 12:123 /1471-2229/12/123 Schiller et al. BMC Plant Biology 2012, 12:123 /1471-2229/12/123 RESEARCH ARTICLE Open Access The Arabidopsis apyrase AtAPY1 is localized in the Golgi instead of the extracellular space 1 1 2 1* Madlen Schiller , Carolin Massalski , Thomas Kurth and Iris Steinebrunner Abstract Background: The two highly similar Arabidopsis apyrases AtAPY1 and AtAPY2 were previously shown to be involved in plant growth and development, evidently by regulating extracellular ATP signals. The subcellular localization of AtAPY1 was investigated to corroborate an extracellular function. Results: Transgenic Arabidopsis lines expressing AtAPY1 fused to the SNAP-(O6-alkylguanine-DNA alkyltransferase)-tag were used for indirect immunofluorescence and AtAPY1 was detected in punctate structures within the cell. The same signal pattern was found in seedlings stably overexpressing AtAPY1-GFP by indirect immunofluorescence and live imaging. In order to identify the nature of the AtAPY1-positive structures, AtAPY1-GFP expressing seedlings were treated with the endocytic marker stain FM4-64 (N-(3-triethylammoniumpropyl)-4-(p-diethylaminophenyl-hexatrienyl)-pyridinium dibromide) and crossed with a transgenic line expressing the trans-Golgi marker Rab E1d. Neither FM4-64 nor Rab E1d co-localized with AtAPY1. However, live imaging of transgenic Arabidopsis lines expressing AtAPY1-GFP and either the fluorescent protein-tagged