Understanding CRY2 interactions for optical control of intracellular signaling.
Understanding CRY2 interactions for optical control of intracellular signaling.
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DOI：
10.1038/s41467-017-00648-8
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发表时间：
2017-09-15
影响因子：
16.6
通讯作者：
Cui B
中科院分区：
文献类型：
--
作者：
Duan L;Hope J;Ong Q;Lou HY;Kim N;McCarthy C;Acero V;Lin MZ;Cui B
拟南芥隐花色素2（cryptochrome 2，CIB 2）可同时发生光依赖的CIB 2-CIB 2同源寡聚化和CIB 2-CIB 1异源二聚化，这两种聚合反应已被广泛应用于光调控细胞内过程。使用β 2-CIB 1相互作用的应用需要最小的β 2同源寡聚化以避免意外的并发症，而使用β 2-CIB 2相互作用的那些应用更喜欢稳健的同源寡聚化。然而，选择β 2相互作用的类型是不可能的，因为β 2相互作用的分子机制是未知的。在这里，我们报告的CIB 2-CIB 1和CIB 2-CIB 2的相互作用是由良好分离的蛋白质界面在CIB 2的两个末端。N-末端电荷对于CIB 2-CIB 1相互作用是关键的。此外，两个C-末端电荷会影响C12同源寡聚化，正电荷促进寡聚化，负电荷抑制寡聚化。通过工程化C-末端电荷，我们分别开发了具有升高或抑制寡聚化的C12高和C12低，我们使用它们来调节Raf/MEK/ERK信号传导水平。这些结果有助于我们理解光诱导的p53 2相互作用的机制，并增强基于p53 2的光遗传系统的可控性。隐花色素2（Cryptochrome 2，CIB 2）可以形成光调控的CIB 2-CIB 2同源寡聚体或CIB 2-CIB 1异源二聚体，但由于缺乏相互作用机制，调控这些相互作用是困难的。在这里，作者确定了促进同源寡聚体的相互作用，并引入突变以产生低和高寡聚化版本。
Arabidopsis cryptochrome 2 (CRY2) can simultaneously undergo light-dependent CRY2–CRY2 homo-oligomerization and CRY2–CIB1 hetero-dimerization, both of which have been widely used to optically control intracellular processes. Applications using CRY2–CIB1 interaction desire minimal CRY2 homo-oligomerization to avoid unintended complications, while those utilizing CRY2–CRY2 interaction prefer robust homo-oligomerization. However, selecting the type of CRY2 interaction has not been possible as the molecular mechanisms underlying CRY2 interactions are unknown. Here we report CRY2–CIB1 and CRY2–CRY2 interactions are governed by well-separated protein interfaces at the two termini of CRY2. N-terminal charges are critical for CRY2–CIB1 interaction. Moreover, two C-terminal charges impact CRY2 homo-oligomerization, with positive charges facilitating oligomerization and negative charges inhibiting it. By engineering C-terminal charges, we develop CRY2high and CRY2low with elevated or suppressed oligomerization respectively, which we use to tune the levels of Raf/MEK/ERK signaling. These results contribute to our understanding of the mechanisms underlying light-induced CRY2 interactions and enhance the controllability of CRY2-based optogenetic systems. Cryptochrome 2 (CRY2) can form light-regulated CRY2-CRY2 homo-oligomers or CRY2-CIB1 hetero-dimers, but modulating these interactions is difficult owing to the lack of interaction mechanism. Here the authors identify the interactions facilitating homo-oligomers and introduce mutations to create low and high oligomerization versions.
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影响因子：
64.8
作者：
通讯作者：
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基于LOV的光遗传设备：光驱动的模块，以赋予细胞信号传导的光心调节控制。
影响因子：
5
作者：
Pudasaini A;El-Arab KK;Zoltowski BD
通讯作者：
Zoltowski BD
影响因子：
4.6
作者：
Katsura Y;Kubota H;Kunida K;Kanno A;Kuroda S;Ozawa T
通讯作者：
Ozawa T
影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
4.7
作者：
Che, Daphne L.;Duan, Liting;Cui, Bianxiao
通讯作者：
Cui, Bianxiao