Benchmarking of optical dimerizer systems.
Benchmarking of optical dimerizer systems.
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DOI：
10.1021/sb500291r
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发表时间：
2014-11-21
影响因子：
4.7
通讯作者：
Tucker, Chandra L.
中科院分区：
文献类型：
--
作者：
Pathak, Gopal P.;Strikland, Devin;Vrana, Justin D.;Tucker, Chandra L.
光学二聚体是一类强大的新型光遗传学工具，其允许光诱导控制蛋白质-蛋白质相互作用。这样的工具已经用于调节细胞通路和过程，在活细胞中具有高时空分辨率，并且越来越多的二聚系统是可用的。由于这些系统的特点是由不同的群体使用不同的方法，用户很难比较它们的属性。在这里，我们开始系统地基准测试四个光学二聚系统，phyB/CIB 1，TULIPs，phyB/PIF 3和phyB/PIF 6的性能。使用酵母转录测定，我们发现红光调节系统之间的光敏感性和倍数激活水平的显着差异，但在TIP 2/CIB和TULIP系统之间的响应相似。进一步比较P172/CIB 1和TULIP系统调节酵母MAPK信号传导途径的能力也显示出类似的反应，用P172/CIB观察到的黑暗中的背景活性略低。在开展这项工作的过程中，我们还在酵母中使用BCL 2/CIB产生了一个改进的蓝光调控转录系统。此外，我们证明了成功的应用程序中的BIP 2/CIB二聚体使用膜栓系的BIP 2，这可能会允许更好地局部控制蛋白质相互作用。总之，这项工作允许更好地理解这些不同的二聚系统的能力，并展示了这些二聚体在酵母中控制信号传导和转录的新用途。
Optical dimerizers are a powerful new class of optogenetic tools that allow light-inducible control of protein–protein interactions. Such tools have been useful for regulating cellular pathways and processes with high spatiotemporal resolution in live cells, and a growing number of dimerizer systems are available. As these systems have been characterized by different groups using different methods, it has been difficult for users to compare their properties. Here, we set about to systematically benchmark the properties of four optical dimerizer systems, CRY2/CIB1, TULIPs, phyB/PIF3, and phyB/PIF6. Using a yeast transcriptional assay, we find significant differences in light sensitivity and fold-activation levels between the red light regulated systems but similar responses between the CRY2/CIB and TULIP systems. Further comparison of the ability of the CRY2/CIB1 and TULIP systems to regulate a yeast MAPK signaling pathway also showed similar responses, with slightly less background activity in the dark observed with CRY2/CIB. In the process of developing this work, we also generated an improved blue-light-regulated transcriptional system using CRY2/CIB in yeast. In addition, we demonstrate successful application of the CRY2/CIB dimerizers using a membrane-tethered CRY2, which may allow for better local control of protein interactions. Taken together, this work allows for a better understanding of the capacities of these different dimerization systems and demonstrates new uses of these dimerizers to control signaling and transcription in yeast.
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DOI：
10.1083/jcb.201210119
发表时间：
2013-05-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Chen D;Gibson ES;Kennedy MJ
通讯作者：
Kennedy MJ
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：
14.9
作者：
Müller K;Engesser R;Schulz S;Steinberg T;Tomakidi P;Weber CC;Ulm R;Timmer J;Zurbriggen MD;Weber W
通讯作者：
Weber W
影响因子：
46.9
作者：
Yazawa, Masayuki;Sadaghiani, Amir M.;Dolmetsch, Ricardo E.
通讯作者：
Dolmetsch, Ricardo E.
影响因子：
3.3
作者：
Pittman, Adrianne M. C.;Lage, Melissa D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.