Illuminating the function of the understudied kinase DYRK2 in ciliary Hedgehog signal transduction

摘要 我们定义了不同生物的纤毛蛋白质组，包括海胆和海葵，以及 确定了DYRK2，一种研究较少的激酶，以前没有涉及到纤毛生物学。后续研究 证实DYRK2定位于纤毛，并揭示DYRK2的缺失破坏了纤毛的形态。我们也 发现DYRK2参与睫状体刺猬的信号转导，沟通沟通 和GLI转录因子，这是该途径的两个中心组成部分。小鼠Dyrk2基因突变导致 骨骼缺陷让人想起那些因失去印度刺猬而造成的缺陷。就像Dyrk2突变一样，药理学上 抑制DYRK2失调的纤毛长度控制和减弱Hedgehog信号。 在这个试点项目中，我们将研究DYRK2在纤毛形态中发挥作用的分子机制， 刺猬信号、骨骼发育和癌症。更具体地说，我们将调查DYRK2是如何运行的 Smoothed下游转导Hedgehog信号(目标1)，DYRK2如何在骨骼中发挥作用 发展(目标2)，以及DYRK2的药理抑制是否可能是一种易治的治疗方法 Hedgehog途径相关癌症(目标3)。拟议的实验使用了一种哺乳动物和 用遗传学、细胞生物学、成像和生化方法揭示DYRK2在睫状刺猬中的功能 信号转导，在发育和疾病中都有。

ABSTRACT We defined the proteomes of cilia from diverse organisms, including sea urchins and sea anemones and identified DYRK2, a poorly studied kinase not been previously implicated in ciliary biology. Subsequent study confirmed that DYRK2 localizes to cilia and revealed that loss of DYRK2 disrupts ciliary morphology. We also found that DYRK2 participates in ciliary Hedgehog signal transduction, communicating between Smoothened and GLI transcription factors, two central components of the pathway. Mutation of mouse Dyrk2 resulted in skeletal defects reminiscent of those caused by loss of Indian hedgehog. Like Dyrk2 mutations, pharmacological inhibition of DYRK2 dysregulated ciliary length control and attenuated Hedgehog signaling. In this pilot project, we will examine the molecular mechanisms by which DYRK2 functions in ciliary morphology, Hedgehog signaling, skeletal development and cancer. More specifically, we will investigate how DYRK2 acts downstream of Smoothened to transduce Hedgehog signals (Aim 1), how DYRK2 functions in skeletal development (Aim 2), and whether pharmacological inhibition of DYRK2 may be a tractable therapy for Hedgehog pathway-associated cancers (Aim 3). The proposed experiments use a combination of mammalian genetic, cell biological, imaging and biochemical approaches to reveal how DYRK2 functions in ciliary Hedgehog signal transduction, both in development and disease.