Kinome siRNA Screen Identifies Regulators of Ciliogenesis and Hedgehog Signal Transduction
Kinome siRNA Screen Identifies Regulators of Ciliogenesis and Hedgehog Signal Transduction
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DOI：
10.1126/scisignal.1162925
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发表时间：
2008-09-30
影响因子：
7.3
通讯作者：
de Sauvage, Frederic J.
中科院分区：
文献类型：
--
作者：
Evangelista, Marie;Lim, Tze Yang;de Sauvage, Frederic J.
Hedgehog (Hh) 通路的破坏或不当激活与发育异常和癌症有关。尽管在果蝇中已得到表征，但在脊椎动物中介导 Smoothened (Smo) 七次跨膜蛋白下游 Hh 信号的机制仍知之甚少。特别是，在果蝇中介导 Hh 信号传导的融合 (Fu) 激酶在哺乳动物中是可有可无的。为了鉴定在哺乳动物中正向调节 Hh 通路的激酶，我们筛选了小鼠激酶组小干扰 RNA 文库，并验证了 9 个调节 Hh 信号传导的候选激酶。在这些候选者中，Nek1 和 Prkra 并不直接在 Hh 通路中发挥作用，而是通过纤毛发生中的主要作用间接对 Hh 信号传导发挥作用。相比之下，另一种激酶 Cdc2l1 直接参与 Hh 通路。 Cdc2l1 对于 Hh 通路的激活是必要且充分的，在 Smo 下游和神经胶质瘤相关 (Gli) 转录因子上游发挥作用。更具体地说，Cdc2l1 与负调节因子 Fused Suppressor (Sufu) 相互作用，解除其对 Gli 的抑制，从而为 Cdc2l1 在 Hh 信号传导中发挥作用提供了一种机制。最后，以斑马鱼为模型生物，我们发现 Cdc2l1 在体内激活 Hh 通路。我们认为 Cdc2l1 是 Hh 信号转导级联中以前未被识别的成员。
Disruption or improper activation of the Hedgehog (Hh) pathway is associated with developmental abnormalities and cancer. Although characterized in Drosophila, the mechanisms that mediate the Hh signal downstream of the Smoothened (Smo) seven-transmembrane protein in vertebrates remain poorly understood. In particular, the Fused (Fu) kinase, which mediates Hh signaling in flies, is dispensable in mammals. To identify kinases that positively regulate the Hh pathway in mammals, we screened a mouse kinome small interfering RNA library and validated nine candidates that modulate Hh signaling. Among these candidates, Nek1 and Prkra did not directly function in the Hh pathway but exerted their effects on Hh signaling indirectly through a primary role in ciliogenesis. In contrast, another kinase, Cdc2l1, directly participated in the Hh pathway. Cdc2l1 was necessary and sufficient for activation of the Hh pathway, functioning downstream of Smo and upstream of the Glioma-associated (Gli) transcription factors. More specifically, Cdc2l1 interacted with the negative regulator Suppressor of Fused (Sufu) and relieved its inhibition on Gli, thus providing a mechanism for how Cdc2l1 might play a role in Hh signaling. Finally, with zebrafish as model organism, we showed that Cdc2l1 activated the Hh pathway in vivo. We propose that Cdc2l1 is a previously unrecognized member of the Hh signal transduction cascade.