Engineered Photoactivatable Genetic Switches Based on the Bacterium Phage T7 RNA Polymerase
Engineered Photoactivatable Genetic Switches Based on the Bacterium Phage T7 RNA Polymerase
复制标题
基于细菌噬菌体 T7 RNA 聚合酶的工程光激活基因开关
DOI：
10.1021/acssynbio.6b00248
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发表时间：
2017-02-01
影响因子：
4.7
通讯作者：
Liu, Haiyan
中科院分区：
文献类型：
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作者：
Han, Tiyun;Chen, Quan;Liu, Haiyan
T7 RNA聚合酶(RNAP)的活性直接受外部信号调控的遗传开关是通过将蛋白质裂解成片段并使用调节结构域来调节它们的重组的工程策略获得的。以光可激活的VVD结构域及其变体作为调节域，获得了具有良好的暗/亮特性的健壮的可切换系统。对于找到的最佳分裂位置，工作开关要么利用VVD域之间的光诱导相互作用，要么利用变构效应。当分裂片段与不同的调控结构域结合时，显示出高度的模块化，例如那些具有化学诱导相互作用的结构域，从而实现化学控制的开关。综上所述，基于T7RNA聚合酶的开关是在不同环境中实现光激活基因表达的强大工具。此外，关于所研究的分裂位置和结构域组织的结果可能有助于未来对该蛋白质和相关蛋白质的工程研究。
Genetic switches in which the activity of T7 RNA polymerase (RNAP) is directly regulated by external Signals are obtained with an engineering strategy of splitting the protein into fragments and using regulatory domains to modulate their reconstitution. Robust switchable systems with excellent dark-off/light-on properties are obtained with the light-activatable VVD domain and its variants as regulatory domains. For the best split position found, working switches exploit either the light-induced interactions between the VVD domains or allosteric effects. The split fragments show high modularity when they are combined with different regulatory domains such as those with chemically inducible interaction, enabling chemically controlled switches. To summarize, the T7 RNA polymerase-based switches are powerful tools to implement light-activated gene expression in different contexts. Moreover, results about the studied split positions and domain organizations may facilitate future engineering studies on this and on related proteins.