Ziomek Krzysztof, Kierzek Elzbieta, Biała Ewa, Kierzek Ryszard
Institute of Bioorganic Chemistry, Polish Academy of Sciences, 60-704 Poznañ, Noskowskiego 12/14, Poland.
Biophys Chem. 2002 Jun 19;97(2-3):233-41. doi: 10.1016/s0301-4622(02)00074-1.
The presence of various modifications within oligomers changes their thermodynamic stability. To get more systematic data, we measured effects of 5- and 6-substituted uridine on thermal stability of (AUCU(Mod.)AGAU)2 and (AUCUAGAU(Mod.))2. Collected results lead to the following conclusions: (i) 5-halogenated and 5-alkylated substituents of the uridine affect thermal stability of the RNA duplexes differently. Moreover, the 5-fluorouridine changes stability of the RNA duplexes opposite to remaining 5-halogenouridines; (ii) for oligomers containing 5-chloro, 5-bromo or 5-iodouridine stronger hydrogen bond formed between oxygen-4 of the 5-halogenated uracil and 6-amino group of the adenine is presumably responsible for stabilizing effect; (iii) placing of A-U(5R) base pairs closer to the end of the duplex enhance thermal stability relatively to oligomer with central position of this base pair; (iv) the effects of 5-substituents are additive, particularly for substituents which stabilize RNA duplexes; (v) 6-methyluridines (N1 and N3 isomers) as well as 3N-methyluridine present at internal position of A-U(Mod.) inhibit duplexes formation; (vi) 6-methyluridines (N1 and N3 isomers) as well as 3N-methyluridine placed as terminal base pairs stabilize the duplexes mostly via 3'-dangling end effect.
寡聚物中各种修饰的存在会改变其热力学稳定性。为了获得更系统的数据,我们测量了5-和6-取代尿苷对(AUCU(Mod.)AGAU)2和(AUCUAGAU(Mod.))2热稳定性的影响。收集到的结果得出以下结论:(i)尿苷的5-卤代和5-烷基取代基对RNA双链体热稳定性的影响不同。此外,5-氟尿苷改变RNA双链体稳定性的方式与其余5-卤代尿苷相反;(ii)对于含有5-氯、5-溴或5-碘尿苷的寡聚物,5-卤代尿嘧啶的氧-4与腺嘌呤的6-氨基之间形成的更强氢键可能是其稳定作用的原因;(iii)相对于该碱基对位于中心位置的寡聚物,将A-U(5R)碱基对放置得更靠近双链体末端会提高热稳定性;(iv)5-取代基的影响是累加的,特别是对于稳定RNA双链体的取代基;(v)位于A-U(Mod.)内部位置的6-甲基尿苷(N1和N3异构体)以及3N-甲基尿苷会抑制双链体的形成;(vi)作为末端碱基对放置的6-甲基尿苷(N1和N3异构体)以及3N-甲基尿苷主要通过3'-悬垂末端效应使双链体稳定。