Distinct Biochemical Activities of Eyes absent During Drosophila Eye Development.
Distinct Biochemical Activities of Eyes absent During Drosophila Eye Development.
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DOI：
10.1038/srep23228
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发表时间：
2016-03-16
影响因子：
4.6
通讯作者：
Mardon G
中科院分区：
文献类型：
--
作者：
Jin M;Mardon G
Eya是一种高度保守的转录辅激活因子和蛋白磷酸酶，在从果蝇到人类的多种发育过程中起着重要作用。Eya蛋白含有富含PST（脯氨酸-丝氨酸-苏氨酸）的反式激活结构域、苏氨酸磷酸酶基序（TPM）和酪氨酸蛋白磷酸酶结构域。使用基因组拯救系统，我们发现PST结构域是必不可少的Eya活性和Dac的表达，和TPM所需的Eya功能。我们还发现，苏氨酸磷酸酶活性在果蝇眼发育过程中只起次要作用，PST和TPM结构域的主要功能是反式激活，可以在很大程度上被异源激活结构域VP 16取代。沿着我们以前的结果，Eya的酪氨酸磷酸酶活性是正常Eya功能的眼睛形成，我们证明了Eya在果蝇眼睛发育的主要功能是作为一个转录辅激活因子。此外，PST/TPM和苏氨酸磷酸酶活性不是视网膜决定因子之间体外相互作用所必需的。最后，这项工作是Eya-Ey物理相互作用的第一份报告。这些发现特别重要，因为它们突出了对准确剖析蛋白质功能的体内方法的需求。
Eyes absent (Eya) is a highly conserved transcriptional coactivator and protein phosphatase that plays vital roles in multiple developmental processes from Drosophila to humans. Eya proteins contain a PST (Proline-Serine-Threonine)-rich transactivation domain, a threonine phosphatase motif (TPM), and a tyrosine protein phosphatase domain. Using a genomic rescue system, we find that the PST domain is essential for Eya activity and Dac expression, and the TPM is required for full Eya function. We also find that the threonine phosphatase activity plays only a minor role during Drosophila eye development and the primary function of the PST and TPM domains is transactivation that can be largely substituted by the heterologous activation domain VP16. Along with our previous results that the tyrosine phosphatase activity of Eya is dispensable for normal Eya function in eye formation, we demonstrate that a primary function of Eya during Drosophila eye development is as a transcriptional coactivator. Moreover, the PST/TPM and the threonine phosphatase activity are not required for in vitro interaction between retinal determination factors. Finally, this work is the first report of an Eya-Ey physical interaction. These findings are particularly important because they highlight the need for an in vivo approach that accurately dissects protein function.