Biophysics, University of Michigan, 930 North University Avenue, Ann Arbor, Michigan 48109-1055, USA.
Acc Chem Res. 2010 Jan 19;43(1):40-7. doi: 10.1021/ar900093g.
Structure and dynamics are both critical to RNA's vital functions in biology. Numerous techniques can elucidate the structural dynamics of RNA, but computational approaches based on experimental data arguably hold the promise of providing the most detail. In this Account, we highlight areas wherein molecular dynamics (MD) and quantum mechanical (QM) techniques are applied to RNA, particularly in relation to complementary experimental studies. We have expanded on atomic-resolution crystal structures of RNAs in functionally relevant states by applying explicit solvent MD simulations to explore their dynamics and conformational changes on the submicrosecond time scale. MD relies on simplified atomistic, pairwise additive interaction potentials (force fields). Because of limited sampling, due to the finite accessible simulation time scale and the approximated force field, high-quality starting structures are required. Despite their imperfection, we find that currently available force fields empower MD to provide meaningful and predictive information on RNA dynamics around a crystallographically defined energy minimum. The performance of force fields can be estimated by precise QM calculations on small model systems. Such calculations agree reasonably well with the Cornell et al. AMBER force field, particularly for stacking and hydrogen-bonding interactions. A final verification of any force field is accomplished by simulations of complex nucleic acid structures. The performance of the Cornell et al. AMBER force field generally corresponds well with and augments experimental data, but one notable exception could be the capping loops of double-helical stems. In addition, the performance of pairwise additive force fields is obviously unsatisfactory for inclusion of divalent cations, because their interactions lead to major polarization and charge-transfer effects neglected by the force field. Neglect of polarization also limits, albeit to a lesser extent, the description accuracy of other contributions, such as interactions with monovalent ions, conformational flexibility of the anionic sugar-phosphate backbone, hydrogen bonding, and solute polarization by solvent. Still, despite limitations, MD simulations are a valid tool for analyzing the structural dynamics of existing experimental structures. Careful analysis of MD simulations can identify problematic aspects of an experimental RNA structure, unveil structural characteristics masked by experimental constraints, reveal functionally significant stochastic fluctuations, evaluate the structural role of base ionization, and predict structurally and potentially functionally important details of the solvent behavior, including the presence of tightly bound water molecules. Moreover, combining classical MD simulations with QM calculations in hybrid QM/MM approaches helps in the assessment of the plausibility of chemical mechanisms of catalytic RNAs (ribozymes). In contrast, the reliable prediction of structure from sequence information is beyond the applicability of MD tools. The ultimate utility of computational studies in understanding RNA function thus requires that the results are neither blindly accepted nor flatly rejected, but rather considered in the context of all available experimental data, with great care given to assessing limitations through the available starting structures, force field approximations, and sampling limitations. The examples given in this Account showcase how the judicious use of basic MD simulations has already served as a powerful tool to help evaluate the role of structural dynamics in biological function of RNA.
结构和动态对于 RNA 在生物学中的重要功能都是至关重要的。许多技术可以阐明 RNA 的结构动态,但基于实验数据的计算方法无疑具有提供最详细信息的潜力。在本专题介绍中,我们强调了分子动力学 (MD) 和量子力学 (QM) 技术在 RNA 中的应用领域,特别是在与互补实验研究相关的领域。我们通过应用显式溶剂 MD 模拟来扩展 RNA 在功能相关状态下的原子分辨率晶体结构,以探索其在亚微秒时间尺度上的动力学和构象变化。MD 依赖于简化的原子间、两两可加相互作用势能(力场)。由于有限的可访问模拟时间尺度和近似的力场,采样受到限制,因此需要高质量的起始结构。尽管存在缺陷,但我们发现,目前可用的力场可以使 MD 能够提供有关 RNA 在晶体学定义的能量最小值周围的动态的有意义且可预测的信息。通过对小模型系统进行精确的 QM 计算可以估计力场的性能。这些计算与 Cornell 等人的 AMBER 力场的结果非常吻合,特别是对于堆积和氢键相互作用。通过模拟复杂的核酸结构可以最终验证任何力场。Cornell 等人的 AMBER 力场的性能通常与实验数据吻合良好并能增强实验数据,但一个值得注意的例外可能是双链茎的帽环。此外,对于二价阳离子的相互作用,两两可加力场的性能显然不能令人满意,因为它们的相互作用会导致力场忽略的主要极化和电荷转移效应。忽略极化也会以较小的程度限制其他相互作用(如与单价离子的相互作用、阴离子糖磷酸骨架的构象灵活性、氢键和溶剂的溶质极化)的描述准确性。尽管存在局限性,MD 模拟仍然是分析现有实验结构的结构动态的有效工具。对 MD 模拟的仔细分析可以识别实验 RNA 结构的问题方面,揭示受实验限制掩盖的结构特征,揭示功能上重要的随机波动,评估碱基离解的结构作用,并预测溶剂行为的结构和潜在功能重要细节,包括紧密结合的水分子的存在。此外,在混合 QM/MM 方法中结合经典 MD 模拟和 QM 计算有助于评估催化 RNA(核酶)的化学机制的合理性。相比之下,根据序列信息可靠地预测结构超出了 MD 工具的适用性。因此,要理解 RNA 功能,计算研究的最终实用性要求既不能盲目接受也不能断然拒绝,而是要结合所有可用的实验数据进行考虑,特别要注意通过可用的起始结构、力场近似和采样限制来评估限制。本专题介绍中的示例展示了明智使用基本 MD 模拟如何已经成为帮助评估结构动态在 RNA 生物学功能中的作用的有力工具。