In vivo regulation of Yorkie phosphorylation and localization
In vivo regulation of Yorkie phosphorylation and localization
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DOI：
10.1242/dev.015255
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发表时间：
2008-03-15
期刊：
影响因子：
4.6
通讯作者：
Irvine, Kenneth D.
中科院分区：
文献类型：
--
作者：
Oh, Hyangyee;Irvine, Kenneth D.
Yorkie（Yki）是Fat和Hippo信号通路的转录因子，受Warts激酶负调控。在这里，我们使用Phos标签凝胶来表征疣依赖的Yki在体内的磷酸化，并显示疣促进Yki在多个位点的磷酸化。我们还表明，疣抑制Yki核定位在体内，并可以促进结合Yki的14-3-3蛋白在培养的细胞。对疣的单个上游调节因子的影响的体内评估揭示了一些突变体（例如，G. fat）对Yki磷酸化仅有部分影响，对Yki定位影响微弱，而其他基因型（e. G. ex脂肪双突变体）对Yki磷酸化和定位都具有更强的影响。我们还确定丝氨酸168作为一个关键的网站，通过Warts介导的磷酸化Yki的负调控发生，但发现这个网站是不足以解释的影响，海马信号Yki在体内。这些结果鉴定了亚细胞定位的调节作为Yki调节的机制，并且确立了这种调节通过Yki上的疣依赖性磷酸化的多个位点在体内发生。
Yorkie (Yki), a transcription factor of the Fat and Hippo signaling pathways, is negatively regulated by the Warts kinase. Here, we use Phos-tag gels to characterize Warts-dependent phosphorylation of Yki in vivo, and show that Warts promotes phosphorylation of Yki at multiple sites. We also show that Warts inhibits Yki nuclear localization in vivo, and can promote binding of Yki to 14-3-3 proteins in cultured cells. In vivo assessment of the influence of individual upstream regulators of Warts reveals that some mutants (e. g. fat) have only partial effects on Yki phosphorylation, and weak effects on Yki localization, whereas other genotypes (e. g. ex fat double mutants) have stronger effects on both Yki phosphorylation and localization. We also identify serine 168 as a critical site through which negative regulation of Yki by Warts-mediated phosphorylation occurs, but find that this site is not sufficient to explain effects of Hippo signaling on Yki in vivo. These results identify modulation of subcellular localization as a mechanism of Yki regulation, and establish that this regulation occurs in vivo through multiple sites of Warts-dependent phosphorylation on Yki.