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KMID : 0613820170270111369
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Journal of Life Science
2017 Volume.27 No. 11 p.1369 ~ p.1375
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Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1
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Kim Na-Young
Kim Moo-Sang
Jung Sung-Hee
Kim Myoung-Sug
Cho Mi-Young
Chung Joon-Ki
Ahn Sang-Jung
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Abstract
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The purpose of this study was to investigate the cellular characterization of phospholipase C-δ1 in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-δ1 (PoPLC-δ1A), which has a high similarity to the human isoform PLC-δ; type-B PoPLC-δ1 (PoPLC-δ1B [Sf]), which has a low similarity to the human isoform PLC-δ and the alternative splice variant PoPLC-δ1B (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-δ1A, PoPLC-δ1B (Sf) and PoPLC-δ1B (Lf). It administered treatments of Ca2+ ionophore ionomycin and endoplasmic reticulum (ER)?Ca2+ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-δ1A was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-δ1B (Lf) and PoPLC-δ1B (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-δ1A in the nuclei when PoPLC-δ1B (Lf) nucleocytoplasmic shuttling and PoPLC-δ1B (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-δ1A, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.
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KEYWORD
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Cell localization, olive flounder, Paralichthys olivaceus, phospholipase C (PLC), translocation
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