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Conformational Heterogeneity in a Fully Complementary DNA Three-Way Junction with a GC-Rich Branchpoint
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-01 00:00:00 , DOI: 10.1021/acs.biochem.7b00677
Anita Toulmin 1, 2 , Laura E. Baltierra-Jasso 1, 2, 3 , Michael J. Morten 3 , Tara Sabir 1, 2 , Peter McGlynn 4 , Gunnar F. Schröder 5, 6 , Brian O. Smith 7 , Steven W. Magennis 3
Affiliation  

DNA three-way junctions (3WJs) are branched structures that serve as important biological intermediates and as components in DNA nanostructures. We recently derived the global structure of a fully complementary 3WJ and found that it contained unpaired bases at the branchpoint, which is consistent with previous observations of branch flexibility and branchpoint reactivity. By combining high-resolution single-molecule Förster resonance energy transfer, molecular modeling, time-resolved ensemble fluorescence spectroscopy, and the first 19F nuclear magnetic resonance observations of fully complementary 3WJs, we now show that the 3WJ structure can adopt multiple distinct conformations depending upon the sequence at the branchpoint. A 3WJ with a GC-rich branchpoint adopts an open conformation with unpaired bases at the branch and at least one additional conformation with an increased number of base interactions at the branchpoint. This structural diversity has implications for branch interactions and processing in vivo and for technological applications.

中文翻译:

具有GC富集分支点的完全互补DNA三通结中的构象异质性

DNA三向连接(3WJ)是分支结构,可作为重要的生物中间体和DNA纳米结构的组成部分。我们最近得出了一个完全互补的3WJ的全局结构,并发现它在分支点处包含不成对的碱基,这与以前对分支灵活性和分支点反应性的观察一致。通过结合高分辨率单分子Förster共振能量转移,分子建模,时间分辨整体荧光光谱和前19种方法对完全互补的3WJ进行F核磁共振观察,我们现在表明3WJ结构可以根据分支点的序列采用多种不同的构象。具有富含GC的分支点的3WJ采用开放构象,在分支处具有不成对的碱基,并且至少一个其他构象在分支点处具有增加的碱基相互作用。这种结构多样性对体内的分支相互作用和加工以及技术应用具有影响。
更新日期:2017-09-04
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