Adornetto G, Porchetta A, Palleschi G, Plaxco K W, Ricci F
Dipartimento di Scienze e Tecnologie Chimiche University of Rome Tor Vergata , Via della Ricerca Scientifica , Rome 00133 , Italy . Email:
Consorzio Interuniversitario Biostrutture e Biosistemi "INBB" , Rome 00136 , Italy.
Chem Sci. 2015 Jul 15;6(7):3692-3696. doi: 10.1039/c5sc00228a. Epub 2015 Mar 10.
Here we explore a general strategy for the rational design of nucleic acid catalysts that can be allosterically activated by specific nucleic-acid binding proteins. To demonstrate this we have combined a catalytic DNAzyme sequence and the consensus sequence recognized by specific transcription factors to create a construct exhibiting two low-energy conformations: a more stable conformation lacking catalytic activity and lacking the transcription factor binding site, and a less stable conformation that is both catalytically active and competent to bind the transcription factor. The presence of the target transcription factor pushes the equilibrium between these states towards the latter conformation, concomitantly activating catalysis. To demonstrate this we have designed and characterized two peroxidase-like DNAzymes whose activities are triggered upon binding either TATA binding protein or the microphthalmia-associated transcription factor. Our approach augments the current tool kit for the allosteric control of DNAzymes and ribozymes and, because transcription factors control many key biological functions, could have important clinical and diagnostic applications.
在这里，我们探索了一种合理设计核酸催化剂的通用策略，该催化剂可被特定核酸结合蛋白变构激活。为了证明这一点，我们将催化性DNA酶序列与特定转录因子识别的共有序列相结合，构建了一个具有两种低能构象的结构：一种更稳定的构象，缺乏催化活性且没有转录因子结合位点；另一种较不稳定的构象，具有催化活性且能够结合转录因子。目标转录因子的存在将这些状态之间的平衡推向后者构象，从而激活催化作用。为了证明这一点，我们设计并表征了两种类过氧化物酶DNA酶，它们的活性在与TATA结合蛋白或小眼相关转录因子结合后被触发。我们的方法扩充了当前用于DNA酶和核酶变构控制的工具包，并且由于转录因子控制许多关键生物学功能，因此可能具有重要的临床和诊断应用。
