Developmental Biology, Saarland University, Building 61, 66421 Homburg, Germany.
Inter-Faculty Institute for Cell Biology Animal Genetics, Auf der Morgenstelle 15, 72076 Tübingen, Germany.
Int J Mol Sci. 2019 Apr 19;20(8):1931. doi: 10.3390/ijms20081931.
The homeodomain-interacting protein kinase (Hipk) is the fly representative of the well-conserved group of HIPKs in vertebrates. It was initially found through its characteristic interactions with homeodomain proteins. Hipk is involved in a variety of important developmental processes, such as the development of the eye or the nervous system. In the present study, we set Hipk and the homeodomain proteins Homeobrain (Hbn), Empty spiracles (Ems), and Muscle segment homeobox (Msh) in an enzyme-substrate relationship. These homeoproteins are transcription factors that function during neurogenesis and are, at least in part, conserved in vertebrates. We reveal a physical interaction between Hipk and the three homeodomain proteins in vivo using bimolecular fluorescence complementation (BiFC). In the course of in vitro phosphorylation analysis and subsequent mutational analysis we mapped several Hipk phosphorylation sites of Hbn, Ems, and Msh. The phosphorylation of Hbn, Ems, and Msh may provide further insight into the function of Hipk during development of the nervous system.
同源结构域相互作用蛋白激酶（Hipk）是脊椎动物中高度保守的 HIPK 家族在果蝇中的代表。它最初是通过其与同源结构域蛋白的特征相互作用而被发现的。Hipk 参与了多种重要的发育过程，如眼睛或神经系统的发育。在本研究中，我们将 Hipk 与同源结构域蛋白 Homeobrain（Hbn）、Empty spiracles（Ems）和 Muscle segment homeobox（Msh）建立了酶-底物关系。这些同源蛋白是在神经发生过程中起作用的转录因子，至少在部分上在脊椎动物中是保守的。我们使用双分子荧光互补（BiFC）在体内揭示了 Hipk 与这三个同源结构域蛋白之间的物理相互作用。在体外磷酸化分析和随后的突变分析过程中，我们对 Hbn、Ems 和 Msh 的几个 Hipk 磷酸化位点进行了定位。Hbn、Ems 和 Msh 的磷酸化可能为 Hipk 在神经系统发育过程中的功能提供进一步的深入了解。
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