The Dual Characteristics of Light-Induced Cryptochrome 2, Homo-oligomerization and Heterodimerization, for Optogenetic Manipulation in Mammalian Cells
The Dual Characteristics of Light-Induced Cryptochrome 2, Homo-oligomerization and Heterodimerization, for Optogenetic Manipulation in Mammalian Cells
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DOI：
10.1021/acssynbio.5b00048
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发表时间：
2015-10-01
影响因子：
4.7
通讯作者：
Cui, Bianxiao
中科院分区：
文献类型：
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作者：
Che, Daphne L.;Duan, Liting;Cui, Bianxiao
光感受器隐花色素2(CRY2)已经成为一种强大的光遗传工具，它可以在光诱导下操纵哺乳动物细胞中的各种信号通路和细胞过程，具有很高的时空精度和易用性。然而，也有研究表明，CRY2在蓝光下的行为是复杂的，因为光激发的CRY2可以通过与其二聚化伙伴CIB1结合而发生同源齐聚和异源二聚。为了更好地了解光诱导的哺乳动物细胞中CRY2的活性，本文系统地研究了不同细胞空间中CRY2的同源齐聚作用，以及CRY2同源齐聚和异二聚化活性是如何相互影响的。定量分析表明，与细胞质形式相比，膜结合的CRY2具有显著增强的齐聚活性。虽然CRY2同源齐聚和CRY2-CIB1异源二聚可以同时发生，但某些CIB1融合蛋白的存在可以抑制CRY2同源齐聚。然而，细胞质CRY2通过与膜结合的CIB1相互作用而募集到膜上，可以显著地加强胞浆CRY2的同源齐聚作用。这些结果有助于理解光诱导的CRY2-CRY2和CRY2-CIB1相互作用系统，并可以作为指导建立利用CRY2的双重光遗传特性来探索细胞过程的新策略。
The photoreceptor cryptochrome 2 (CRY2) has become a powerful optogenetic tool that allows light-inducible manipulation of various signaling pathways and cellular processes in mammalian cells with high spatiotemporal precision and ease of application. However, it has also been shown that the behavior of CRY2 under blue light is complex, as the photoexcited CRY2 can both undergo homo-oligomerization and heterodimerization by binding to its dimerization partner CIB1 To better understand the light-induced CRY2 activities in mammalian cells, this article systematically characterizes CRY2 homo-oligomerization in different cellular compartments, as well as how CRY2 homo-oligomerization and heterodimerization activities affect each other. Quantitative analysis reveals that membrane-bound CRY2 has drastically enhanced oligomerization activity compared to that of its cytoplasmic form. While CRY2 homo-oligomerization and CRY2-CIB1 heterodimerization could happen concomitantly, the presence of certain CIB1 fusion proteins can suppress CRY2 homo-oligomerization. However, the homo-oligomerization of cytoplasmic CRY2 can be significantly intensified by its recruitment to the membrane via interaction with the membrane-bound CIB1. These results contribute to the understanding of the light-inducible CRY2-CRY2 and CRY2-CIB1 interaction systems and can be used as a guide to establish new strategies utilizing the dual optogenetic characteristics of CRY2 to probe cellular processes.