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http://dx.doi.org/10.5352/JLS.2017.27.11.1369

Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1  

Kim, Na Young (Pathology Research Division, National Institute of Fisheries Science (NIFS))
Kim, Moo-Sang (Department of Aquatic Life Medicine, Pukyong National University)
Jung, Sung Hee (Pathology Research Division, National Institute of Fisheries Science (NIFS))
Kim, Myoung Sug (Pathology Research Division, National Institute of Fisheries Science (NIFS))
Cho, Mi Young (Pathology Research Division, National Institute of Fisheries Science (NIFS))
Chung, oon Ki (Department of Aquatic Life Medicine, Pukyong National University)
Ahn, Sang Jung (Fisheries R&D Management Center, Korea Institute of Marine Science & Technology Promotion (KIMST))
Publication Information
Journal of Life Science / v.27, no.11, 2017 , pp. 1369-1375 More about this Journal
Abstract
The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ 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-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (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-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.
Keywords
Cell localization; olive flounder; Paralichthys olivaceus; phospholipase C (PLC); translocation;
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