Optogenetic clustering and membrane translocation of the BcLOV4 photoreceptor.
Optogenetic clustering and membrane translocation of the BcLOV4 photoreceptor.
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DOI：
10.1073/pnas.2221615120
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发表时间：
2023-08-08
影响因子：
11.1
通讯作者：
Bugaj, Lukasz J.
中科院分区：
文献类型：
--
作者：
Pal, Ayush Aditya;Benman, William;Mumford, Thomas R.;Huang, Zikang;Chow, Brian Y.;Bugaj, Lukasz J.
光诱导蛋白对于细胞在空间和时间上的精确控制产生了巨大的影响。我们在 BcLOV4 光感受器中发现了一种新型光诱导行为，它同时聚集并易位到哺乳动物细胞的细胞膜上。这种独特的组合为新型光遗传学工具打开了大门。我们还发现聚类和膜易位是相关的，因此增强聚类会增强膜易位。这使我们能够快速生成更有效、更灵敏的基于 BcLOV 的光遗传学探针变体。由于 BcLOV4 在酵母、果蝇、斑马鱼、海鞘和人类等多种生物体的细胞中发挥作用，因此我们的研究结果可以广泛应用。光遗传学工具通过少数行为之一对光作出反应，包括变构变化、二聚化、聚集或膜易位。在这里，我们描述了一种新型光遗传学致动器，它响应蓝光而同时聚集并易位到质膜。我们证明，BcLOV4 光感受器的双重易位和聚类可用于新型单组分光遗传学工具，包括控制整个表皮生长因子受体 (ErbB1-4) 酪氨酸激酶家族。我们进一步发现聚类和膜易位在机制上是相关的。更强的聚类增加了易位和下游信号传导的幅度，对光的敏感性增加了约三到四倍，并降低了强信号激活所需的表达水平。因此，BcLOV4 的光诱导聚类提供了一种生成新型光遗传学工具并增强现有工具的策略。
Light-inducible proteins have had tremendous impact for the precise control of cells in space and time. We found a new type of light-induced behavior in the BcLOV4 photoreceptor, which simultaneously clusters and translocates to the membrane in mammalian cells. This unique combination opens the door to a new class of optogenetic tools. We also find that clustering and membrane translocation are linked such that enhancing clustering enhances membrane translocation. This allows us to rapidly generate more potent and sensitive variants of BcLOV-based optogenetic probes. Because BcLOV4 works in cells from diverse organisms including yeast, flies, zebrafish, ciona, and humans, our findings can be applied broadly. Optogenetic tools respond to light through one of a small number of behaviors including allosteric changes, dimerization, clustering, or membrane translocation. Here, we describe a new class of optogenetic actuator that simultaneously clusters and translocates to the plasma membrane in response to blue light. We demonstrate that dual translocation and clustering of the BcLOV4 photoreceptor can be harnessed for novel single-component optogenetic tools, including for control of the entire family of epidermal growth factor receptor (ErbB1-4) tyrosine kinases. We further find that clustering and membrane translocation are mechanistically linked. Stronger clustering increased the magnitude of translocation and downstream signaling, increased sensitivity to light by ~threefold-to-fourfold, and decreased the expression levels needed for strong signal activation. Thus light-induced clustering of BcLOV4 provides a strategy to generate a new class of optogenetic tools and to enhance existing ones.
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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：
Duan, Liting;Hope, Jen M.;Cui, Bianxiao
通讯作者：
Cui, Bianxiao
影响因子：
7.7
作者：
Huang, Yongjian;Bharill, Shashank;Kuriyan, John
通讯作者：
Kuriyan, John
DOI：
10.1073/pnas.91.17.8132
发表时间：
1994-08-16
影响因子：
11.1
作者：
GUY, PM;PLATKO, JV;CARRAWAY, KL
通讯作者：
CARRAWAY, KL
影响因子：
4.8
作者：
Bublil, Erez M.;Pines, Gur;Yarden, Yosef
通讯作者：
Yarden, Yosef