Chalikian T V, Völker J, Srinivasan A R, Olson W K, Breslauer K J
Department of Pharmaceutical Sciences, Faculty of Pharmacy, University of Toronto, 19 Russell Street, Toronto, Ontario, Canada M5S 2S2.
Biopolymers. 1999 Oct 15;50(5):459-71. doi: 10.1002/(SICI)1097-0282(19991015)50:5<459::AID-BIP1>3.0.CO;2-B.
Using high precision densimetric and ultrasonic measurements, we have determined, at 25 degrees C, the apparent molar volumes PhiV and the apparent molar compressibilities PhiK(S) of four nucleic acid duplexes-namely, the DNA duplex, poly(dIdC)poly(dIdC); the RNA duplex, poly(rA)poly(rU); and the two DNA/RNA hybrid duplexes, poly(rA)poly(dT) and poly(dA)poly(rU). Using available fiber diffraction data on these duplexes, we have calculated the molecular volumes as well as the solvent-accessible surface areas of the constituent charged, polar, and nonpolar atomic groups. We found that the hydration properties of these nucleic acid duplexes do not correlate with the extent and the chemical nature of the solvent-exposed surfaces, thereby suggesting a more specific set of duplex-water interactions beyond general solvation effects. A comparative analysis of our volumetric data on the four duplexes, in conjunction with available structural information, suggests the following features of duplex hydration: (a) The four duplexes exhibit different degrees of hydration, in the order poly(dIdC)poly(dIdC) > poly(dGdC)poly(dGdC) > poly(dAdT)poly(dAdT) approximately poly(dA)poly(dT). (b) Repetitive AT and IC sequences within a duplex are solvated beyond general effects by a spine of hydration in the minor groove, with this sequence-specific water network involving about 8 additional water molecules from the second and, perhaps, even the third hydration layers. (c) Repetitive GC and IC sequences within a duplex are solvated beyond general effects by a "patch of hydration" in the major groove, with this water network involving about 13 additional water molecules from the second and, perhaps, even the third hydration layers. (d) Random sequence, polymeric DNA duplexes, which statistically lack extended regions of repetitive AT, GC, or IC sequences, do not experience such specific enhancements of hydration. Consequently, consistent with our previous observations (T. V. Chalikian, A. P. Sarvazyan, G. E. Plum, and K. J. Breslauer, Biochemistry, 1994, Vol. 33, pp. 2394-2401), duplexes with approximately 50% AT content exhibit the weakest hydration, while an increase or decrease from this AT content causes enhancement of hydration, either due to stronger hydration of the minor groove (an increase in AT content) or due to stronger hydration of the major groove (an increase in GC content). (e) In dilute aqueous solutions, a B-DNA duplex is more hydrated than an A-DNA duplex, a volumetric-based conclusion that is in agreement with previous results obtained on crystals, fibers, and DNA solutions in organic solvent-water mixtures. (f) the A-like, RNA duplex poly(rA)poly(rU) and the structurally similar A-like, hybrid duplex poly(rA)poly(dT), exhibit similar hydration properties, while the structurally distinct A-like, hybrid duplex poly(rA)poly(dT) and non-A-like, hybrid duplex poly(dA)poly(rU) exhibit differential hydration properties, consistent with structural features dictating hydration characteristics. We discuss how volumetric characterizations, in conjunction with structural studies, can be used to describe, define, and resolve the general and sequence/conformation-specific hydration properties of nucleic acid duplexes.
通过高精度密度测定和超声测量,我们在25摄氏度下测定了四种核酸双链体的表观摩尔体积ΦV和表观摩尔压缩系数ΦK(S),这四种双链体分别是:DNA双链体聚(dIdC)聚(dIdC);RNA双链体聚(rA)聚(rU);以及两种DNA/RNA杂交双链体聚(rA)聚(dT)和聚(dA)聚(rU)。利用这些双链体现有的纤维衍射数据,我们计算了组成双链体的带电、极性和非极性原子基团的分子体积以及溶剂可及表面积。我们发现,这些核酸双链体的水合特性与溶剂暴露表面的范围和化学性质无关,从而表明除了一般的溶剂化作用外,还存在一组更特殊的双链体 - 水相互作用。对这四种双链体的体积数据进行的比较分析,结合现有的结构信息,揭示了双链体水合的以下特征:(a)这四种双链体表现出不同程度的水合作用,顺序为聚(dIdC)聚(dIdC)>聚(dGdC)聚(dGdC)>聚(dAdT)聚(dAdT)≈聚(dA)聚(dT)。(b)双链体内的重复AT和IC序列通过小沟中的水合脊进行溶剂化,超出了一般效应,这种序列特异性水网络涉及来自第二甚至第三水合层的约8个额外水分子。(c)双链体内的重复GC和IC序列通过大沟中的“水合斑块”进行溶剂化,超出了一般效应,这种水网络涉及来自第二甚至第三水合层的约13个额外水分子。(d)随机序列的聚合DNA双链体,在统计学上缺乏重复AT、GC或IC序列的延伸区域,不会经历这种水合作用的特异性增强。因此,与我们之前的观察结果一致(T. V. Chalikian、A. P. Sarvazyan、G. E. Plum和K. J. Breslauer,《生物化学》,1994年,第33卷,第2394 - 2401页),AT含量约为50%的双链体表现出最弱的水合作用,而AT含量从这个值增加或减少都会导致水合作用增强,这要么是由于小沟水合作用增强(AT含量增加),要么是由于大沟水合作用增强(GC含量增加)。(e)在稀水溶液中,B - DNA双链体比A - DNA双链体水合程度更高,这一基于体积的结论与之前在晶体、纤维以及有机溶剂 - 水混合物中的DNA溶液上获得的结果一致。(f)类似A的RNA双链体聚(rA)聚(rU)和结构相似的类似A的杂交双链体聚(rA)聚(dT)表现出相似的水合特性,而结构不同的类似A的杂交双链体聚(rA)聚(dT)和非A样杂交双链体聚(dA)聚(rU)表现出不同的水合特性,这与决定水合特征的结构特征一致。我们讨论了如何将体积表征与结构研究相结合,以描述、定义和解析核酸双链体的一般和序列/构象特异性水合特性。