KMID : 1161520170210050316
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Animal Cells and Systems 2017 Volume.21 No. 5 p.316 ~ p.322
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PI4KII activity-dependent Golgi complex targeting of Aplysia phosphodiesterase 4 long-form mutant
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Jun Yong-Woo
Lee Jin-A Kaang Bong-Kiun Jang Deok-Jin
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Abstract
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The compartmentalization of cAMP by specifically targeted phosphodiesterases (PDEs) contributes to signal regulation in defined regions of cells. We previously demonstrated that the 20 N-terminal amino acids of Aplysia PDE4 (ApPDE4) long-form (L(N20)) and the two mutants of L(N20) were localized to the Golgi complex. However, the molecular mechanisms underlying the Golgi complex targeting of ApPDE4 long-form and its mutated forms are not clear. In the present study, we show that the Golgi complex targeting of L(N20/C14,15S)-enhanced green fluorescent protein (EGFP) was antimycin A-, phenylarsine oxide (PAO)-, and adenosine-sensitive, but insensitive to high concentrations of wortmannin. On the other hand, the Golgi complex targeting of L(N20)-EGFP and L(N20/C3,14S)-EGFP was antimycin A- and PAO-insensitive. These results suggest that the Golgi-localized lipid kinase protein, phosphatidylinositol 4-kinase type II alpha (PI4KII¥á), the activity of which is inhibited by PAO and adenosine, but not by high concentrations of wortmannin, is likely involved in the Golgi complex targeting of L(N20/C14,15S)-EGFP. In addition, subcellular localization of L(N20/C14,15S)-EGFP, but not L(N20)-EGFP or L(N20/C3,14S)-EGFP, was changed from the Golgi complex only to both the endoplasmic reticulum (ER) and the Golgi complex following treatment with a palmitoylation inhibitor, 2-bromo palmitate. Taken together, our results suggest that L(N20/C14,15S)-EGFP, but not L(N20)-EGFP or L(N20/C3,14S)-EGFP, is localized to the Golgi complex in a PI4KII activity- and palmitoylation-dependent manner. Therefore, phosphatidylinositol 4-phosphate (PI4P) generated by PI4KII¥á at the Golgi complex might play a key role in the Golgi complex targeting of L(N20/C14,15S)-EGFP.
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KEYWORD
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Aplysia, phosphodiesterase 4, Golgi complex, phosphatidylinositol 4-phosphate, phosphatidylinositol 4-kinase type II alpha
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