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Casein kinase II phosphorylation regulates alphaNAC subcellular localization and transcriptional coactivating activity.

机译:酪蛋白激酶II磷酸化调节alphaNAC亚细胞定位和转录共激活活性。

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摘要

The subcellular localization of the alphaNAC coactivator is regulated, but the signaling pathways controlling its nucleocytoplasmic shuttling and coactivation function are not completely characterized. We report here that casein kinase II (CK2) phosphorylated alphaNAC on several phosphoacceptor sites, especially in an amino-terminal cluster. Deletion or mutation of the clustered CK2 sites induced nuclear accumulation of alphaNAC in cells. alphaNAC also localized to the nucleus when endogenous CK2 activity was inhibited by quercetin or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). These observations suggested that phosphorylation by CK2 might play a signaling role in the nuclear export of alphaNAC. Interestingly, inhibition of the chromosome region maintenance 1 (CRM1) exportin by leptomycin B (LMB) led to accumulation of alphaNAC in the nucleus. We conclude that CK2 phosphorylation of the N-terminal cluster corresponds to the signal for alphaNAC's nuclear export via a CRM1-dependent pathway. Finally, the nuclear accumulation of the protein resulting from the lack of CK2 phosphorylation mediated a slight but significant increase of the alphaNAC coactivating function on AP-1 transcriptional activity. Thus, alphaNAC's exit from the nucleus and capacity to potentiate transcription appear dependent on its phosphorylation status.
机译:alphaNAC共激活子的亚细胞定位受到调节,但控制其核质穿梭和共激活功能的信号传导途径尚未完全表征。我们在这里报告酪蛋白激酶II(CK2)磷酸化alphaNAC在几个磷酸受体位点,尤其是在氨基末端簇。聚集的CK2位点的缺失或突变诱导了αNAC在细胞中的核积累。当槲皮素或5,6-二氯-1-β-D-呋喃核糖基苯并咪唑(DRB)抑制内源性CK2活性时,alphaNAC也定位于细胞核。这些观察结果表明CK2的磷酸化可能在αNAC的核输出中起信号作用。有趣的是,瘦素B(LMB)对染色体区域维持1(CRM1)输出蛋白的抑制导致alphaNAC在细胞核中积聚。我们得出的结论是,N末端簇的CK2磷酸化对应于alphaNAC通过CRM1依赖性途径的核输出信号。最后,由于缺乏CK2磷酸化而导致蛋白质的核积累介导了对AP-1转录活性的alphaNAC共激活功能的轻微但显着增加。因此,αNAC从细胞核的退出和增强转录的能力似乎取决于其磷酸化状态。

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