open access

https://hdl.handle.net/2115/42566

Visualization of Phosphoinositides via the Development of the Transient Expression System of a Cyan Fluorescent Protein in the Red Alga Porphyra yezoensis

Expression of PH domain-CFP fusions in a red alga

英語

Porphyra yezoensis

2009-10

Springer

Marine Biotechnology

11(5)

563-569

Phosphoinositides (PIs) play important roles in signal transduction pathways and the regulation of cytoskeleton and membrane functions in eukaryotes. Subcellular localization of individual PI derivative is successfully visualized in yeast, animal and green plant cells using PI derivative-specific pleckstrin homology (PH) domains fused with a variety of fluorescent proteins, however expression of fluorescent proteins has not yet been reported in any red algal cells. In the present study, we developed the system to visualize these PIs using human PH domains fused with a humanized cyan fluorescent protein (AmCFP) in the red alga Porphyra yezoensis. Plasma membrane-localization of AmCFP fused with the PH domain from phospholipase Cδ1 and Akt1, but not Bruton's tyrosine kinase, was observed in cell wall-free monospores, demonstrating the presence of phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4-bisphosphate in P. yezoensis cells. This is the first report of the successful expression of fluorescent protein and the monitoring of PI derivatives in red algal cells. Our system based on transient expression of AmCFP could be applicable for the analysis of subcellular localization of other proteins in P. yezoensis and other red algal cells.

学術雑誌論文

Version of Record

The original publication is available at springerlink.com

1436-2228

1436-2236

AA11357643