CRISPR Screens Uncover Genes that Regulate Target Cell Sensitivity to the Morphogen Sonic Hedgehog.
CRISPR Screens Uncover Genes that Regulate Target Cell Sensitivity to the Morphogen Sonic Hedgehog.
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DOI：
10.1016/j.devcel.2017.12.003
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发表时间：
2018-01-08
影响因子：
11.8
通讯作者：
Rohatgi R
中科院分区：
文献类型：
--
作者：
Pusapati GV;Kong JH;Patel BB;Krishnan A;Sagner A;Kinnebrew M;Briscoe J;Aravind L;Rohatgi R
为了揭示Hedgehog （Hh）信号传导的调控机制，我们进行了全基因组筛选，以确定阳性和阴性通路成分，并在两种不同细胞类型中使用多种信号传导和分化试验验证了顶部命中。在我们的筛选中发现的大多数阳性调节因子，包括Rab34、Pdcl和Tubd1，都参与纤毛功能，证实了初级纤毛在Hh信号传导中的核心作用。阴性调节因子包括Megf8、Mgrn1和一个未加注释的基因，我们将其命名为Atthog。这些负调节因子的功能集中在Smoothened （SMO）上，SMO是一种通过细胞膜传导Hh信号的癌蛋白。在缺乏Atthog的情况下，SMO在细胞表面稳定并集中在纤毛膜上，增强细胞对配体Sonic Hedgehog （SHH）的敏感性，从而改变SHH引导的神经细胞命运决定。因此，我们发现了通过调节靶细胞中的蛋白质转运事件来改变形态因子信号解释的基因。CRISPR筛选确定了与纤毛和纤毛病相关的已知和新基因Atthog， Megf8和Mgrn1减弱了靶细胞对Hedgehog信号的反应信号的衰减因子促进Smoothened的内吞和降解Pusapati等人在哺乳动物细胞中使用基于CRISPR的筛选来鉴定修饰细胞对Hedgehog形态因子反应的基因。这些筛选提供了刺猬信号的调控结构的全面视图，突出了其与原发性纤毛和纤毛病的密切联系，并揭示了Smoothened贩运在调节信号强度中的作用。
To uncover regulatory mechanisms in Hedgehog (Hh) signaling, we conducted genome-wide screens to identify positive and negative pathway components and validated top hits using multiple signaling and differentiation assays in two different cell types. Most positive regulators identified in our screens, including Rab34, Pdcl, and Tubd1, were involved in ciliary functions, confirming the central role for primary cilia in Hh signaling. Negative regulators identified included Megf8, Mgrn1, and an unannotated gene encoding a tetraspan protein we named Atthog. The function of these negative regulators converged on Smoothened (SMO), an oncoprotein that transduces the Hh signal across the membrane. In the absence of Atthog, SMO was stabilized at the cell surface and concentrated in the ciliary membrane, boosting cell sensitivity to the ligand Sonic Hedgehog (SHH) and consequently altering SHH-guided neural cell-fate decisions. Thus, we uncovered genes that modify the interpretation of morphogen signals by regulating protein-trafficking events in target cells. CRISPR screens identify positive and negative regulators of Hedgehog signaling Screens identified known and new genes linked to cilia and ciliopathies Atthog, Megf8, and Mgrn1 attenuate target cell responses to Hedgehog signals Attenuators of signaling promote the endocytosis and degradation of Smoothened Pusapati et al. use CRISPR-based screens in mammalian cells to identify genes that modify cellular responses to Hedgehog morphogens. The screens provide a comprehensive view of the regulatory structure of Hedgehog signaling, highlight its intimate connection to primary cilia and ciliopathies, and uncover a role for Smoothened trafficking in regulating signaling strength.
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影响因子：
64.8
作者：
Eggenschwiler, JT;Espinoza, E;Anderson, KV
通讯作者：
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影响因子：
4.3
作者：
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通讯作者：
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影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
14.9
作者：
Lassmann T;Frings O;Sonnhammer EL
通讯作者：
Sonnhammer EL