Real-Time Monitoring of the Multistate Conformational Dynamics of Polypurine Reverse Hoogsteen Hairpin To Capture Their Triplex-Affinity for Gene Silencing by smFRET Microspectroscopy.,The Journal of Physical Chemistry B

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Real-Time Monitoring of the Multistate Conformational Dynamics of Polypurine Reverse Hoogsteen Hairpin To Capture Their Triplex-Affinity for Gene Silencing by smFRET Microspectroscopy.
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2020-08-25 , DOI: 10.1021/acs.jpcb.0c05493
Debolina Bandyopadhyay ₁ , Padmaja P Mishra ₁

Affiliation

Single-molecule Förster resonance energy transfer microspectroscopic methods are employed for real-time monitoring and to gain deeper insights into the formation of the polypurine reverse Hoogsteen hairpin (PPRH) and its triplex-forming activity. The heterogeneity in the behavior of individual PPRHs has been documented, and it is seen that the degree of anharmonic plasticity of the antiparallel hairpin is stabilized by the formation of reverse Hoogsteen (RH) bonds. While being involved in the hairpin formation, they flip reversibly between the open and closed conformations, irrespective of the concentration of ions present in their microenvironment. However, the nature of the cation present in the buffer plays a crucial role in determining the structural stability. The Watson Crick (WC) bonds are found to be more dynamic in the triplex compared to that of the RH base pairs, indicating the involvement of progressive WC bonds during the triplex motif formation by the PPRH. The majority of the intact triplex DNA attained a semifolded relaxed state before progressing toward a tightly folded state, emphasizing the fact that the folding mechanism pursues an ambiguous path in the mode of acquiring the final step of the triple helix motif. Moreover, the presence of triplex-forming sequences in the regulatory regions of the genome further provides an intricate link between the experimental results and sequence occurrence.

中文翻译：

实时监测多嘌呤反向Hougsteen发夹的多态构象动力学，以捕获它们的三链亲和力，从而通过smFRET显微技术实现基因沉默。

单分子Förster共振能量转移显微光谱法用于实时监测，并深入了解多嘌呤反向Hoogsteen发夹（PPRH）的形成及其三链体形成活性。已记录了单个PPRH行为的异质性，并且可以看到反平行发夹的非谐可塑性程度通过反向Hoogsteen（RH）键的形成得以稳定。尽管它们参与发夹结构的形成，但无论其微环境中存在的离子浓度如何，它们都可在开放和闭合构象之间可逆翻转。但是，存在于缓冲液中的阳离子的性质在决定结构稳定性方面起着至关重要的作用。与RH碱基对相比，发现Watson Crick（WC）键在三链体中更具动态性，表明PPRH在三链体基序形成过程中涉及了进行性WC键。完整的三链DNA大部分在进入紧密折叠状态之前已达到半折叠的松弛状态，强调了折叠机制以获取三螺旋基序的最终步骤的方式追求歧义的事实。此外，基因组调控区中三链体形成序列的存在进一步提供了实验结果与序列出现之间的复杂联系。完整的三链DNA大部分在进入紧密折叠状态之前已达到半折叠的松弛状态，强调了折叠机制以获取三螺旋基序的最终步骤的方式追求歧义的事实。此外，基因组调控区中三链体形成序列的存在进一步提供了实验结果与序列出现之间的复杂联系。完整的三链DNA大部分在进入紧密折叠状态之前已达到半折叠的松弛状态，强调了折叠机制以获取三螺旋基序的最终步骤的方式追求歧义的事实。此外，基因组调控区中三链体形成序列的存在进一步提供了实验结果与序列出现之间的复杂联系。

更新日期：2020-09-24

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