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

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Title: Visualization of Phosphoinositides via the Development of the Transient Expression System of a Cyan Fluorescent Protein in the Red Alga Porphyra yezoensis
Other Titles: Expression of PH domain-CFP fusions in a red alga
Authors: Mikami, Koji Browse this author →KAKEN DB
Uji, Toshiki Browse this author →KAKEN DB
Li, Lin Browse this author
Takahashi, Megumu Browse this author
Yasui, Hajime Browse this author →KAKEN DB
Saga, Naotsune Browse this author →KAKEN DB
Keywords: Pleckstrin homology domain
Phosphoinositide
Cyan fluorescent protein
Transient gene expression
Porphyra yezoensis
Issue Date: Oct-2009
Publisher: Springer
Journal Title: Marine Biotechnology
Volume: 11
Issue: 5
Start Page: 563
End Page: 569
Publisher DOI: 10.1007/s10126-008-9172-z
PMID: 19153794
Abstract: 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.
Rights: The original publication is available at springerlink.com
Type: article
URI: http://hdl.handle.net/2115/42566
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 三上 浩司

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