NMR solution structure of the RED subdomain of the Sleeping Beauty transposase.
NMR solution structure of the RED subdomain of the Sleeping Beauty transposase.
复制标题
睡美人转座酶 RED 子结构域的 NMR 溶液结构。
DOI：
10.1002/pro.3167
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Nesmelova,IrinaV
中科院分区：
文献类型：
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作者：
Konnova,TatianaA;Singer,ChristopherM;Nesmelova,IrinaV
DNA转座子可用于脊椎动物中的稳定基因转移。睡美人（Sleeping Beauty，SB）DNA转座子最近已被应用于人类，并正在进行临床试验，但其分子机制尚不清楚。SB转座由转座酶催化，转座酶是一种含有催化结构域和DNA结合结构域的多结构域蛋白。SB转座酶的DNA结合结构域包含两个结构独立的亚结构域，派和RED。最近，催化结构域和派亚结构域的结构已被确定，但没有关于RED亚结构域及其与DNA相互作用的结构信息。在这里，我们使用NMR光谱来确定RED子域的溶液结构，并表征其与转座子DNA的相互作用。
DNA transposons can be employed for stable gene transfer in vertebrates. The Sleeping Beauty (SB) DNA transposon has been recently adapted for human application and is being evaluated in clinical trials, however its molecular mechanism is not clear. SB transposition is catalyzed by the transposase enzyme, which is a multi‐domain protein containing the catalytic and the DNA‐binding domains. The DNA‐binding domain of the SB transposase contains two structurally independent subdomains, PAI and RED. Recently, the structures of the catalytic domain and the PAI subdomain have been determined, however no structural information on the RED subdomain and its interactions with DNA has been available. Here, we used NMR spectroscopy to determine the solution structure of the RED subdomain and characterize its interactions with the transposon DNA.