Light-induced, spatiotemporal control of protein in the developing embryo of the sea urchin.
Light-induced, spatiotemporal control of protein in the developing embryo of the sea urchin.
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DOI：
10.1016/j.ydbio.2021.06.006
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发表时间：
2021-10
影响因子：
2.7
通讯作者：
Yajima M
中科院分区：
文献类型：
--
作者：
Wavreil FDM;Poon J;Wessel GM;Yajima M
差异蛋白调节是调节细胞活性和控制细胞命运决定的关键生物学过程。这是特别重要的早期胚胎发生时，转录后事件占主导地位的差异命运规范在许多生物体。光诱导方法已经成为一种强大的技术，通过在共聚焦显微镜上控制激光照射来询问具有时间和空间精度的蛋白质功能，甚至在细胞内的亚细胞水平上。然而，由于每个系统的复杂性质，这些工具的应用和功效需要针对每个模型系统或针对感兴趣的细胞类型进行测试。在这里，我们介绍了两种类型的光诱导的方法来跟踪和控制蛋白质在亚细胞水平的海胆胚胎发育。我们发现，光转换荧光蛋白，Kaede，是非常有效的区分预先存在的和新合成的蛋白质，没有明显的光毒性，即使与有丝分裂纺锤体相关的蛋白质的询问。此外，使用miniSOG的发色团辅助光灭活（卡利）成功地灭活了植物皮层的感兴趣的蛋白质，并选择性地延迟或抑制了不对称细胞分裂。总的来说，这些光诱导的方法作为额外的分子工具来识别蛋白质活性，不仅在每个细胞谱系中，而且在发育胚胎的亚细胞水平上。
Differential protein regulation is a critical biological process that regulates cellular activity and controls cell fate determination. It is especially important during early embryogenesis when post-transcriptional events predominate differential fate specification in many organisms. Light-induced approaches have been a powerful technology to interrogate protein functions with temporal and spatial precision, even at subcellular levels within a cell by controlling laser irradiation on the confocal microscope. However, application and efficacy of these tools need to be tested for each model system or for the cell type of interest because of the complex nature of each system. Here, we introduce two types of light-induced approaches to track and control proteins at a subcellular level in the developing embryo of the sea urchin. We found that the photoconvertible fluorescent protein, Kaede, is highly efficient to distinguish pre-existing and newly synthesized proteins with no apparent phototoxicity, even with interrogating proteins associated with mitotic spindle. Further, chromophore-assisted light inactivation (CALI) using miniSOG successfully inactivated proteins of interest at the vegetal cortex and selectively delayed or inhibited asymmetric cell division. Overall, these light-induced approaches serve as additional molecular tools to identify protein activity, not only in each cell lineage but also at a subcellular level in developing embryos.
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DOI：
10.1083/jcb.200907090
发表时间：
2009-12-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
von Dassow G;Verbrugghe KJ;Miller AL;Sider JR;Bement WM
通讯作者：
Bement WM
影响因子：
16
作者：
Mizuno, H;Mal, TK;Miyawakil, A
通讯作者：
Miyawakil, A
DOI：
10.1073/pnas.1018513108
发表时间：
2011-05-31
影响因子：
11.1
作者：
Wei, Zheng;Angerer, Robert C.;Angerer, Lynne M.
通讯作者：
Angerer, Lynne M.
影响因子：
21.3
作者：
Miller, Ann L.;Bement, William M.
通讯作者：
Bement, William M.
影响因子：
21.3
作者：
Rajfur, Z;Roy, P;Jacobson, K
通讯作者：
Jacobson, K