Peterson R D, Bartel D P, Szostak J W, Horvath S J, Feigon J
Department of Chemistry, University of California, Los Angeles 90024.
Biochemistry. 1994 May 10;33(18):5357-66. doi: 10.1021/bi00184a001.
1H NMR studies of a 30-nucleotide RNA oligonucleotide (RBE3), which contains a high-affinity binding site for Rev of the HIV-1 Rev responsive element (RRE), two derivatives of RBE3 (RBE3AA and RBE3-A), and the complex of RBE3 with peptides derived from the RNA binding domain of HIV-1 Rev, are presented. The high-affinity binding site of the RRE consists of an asymmetric internal loop and surrounding Watson-Crick base pairs. In the wild-type RRE, one of the stems is closed by a loop; this is replaced in REB3 by the stable UUCG tetraloop. NOE data suggest that the internal loop of the free RNA contains structural features that have been predicted on the basis of in vitro selection experiments [Bartel, D.P., et al. (1991) Cell 67, 529-536]. The structural features include a Gsyn.Ganti base pair, a Ganti.Aanti base pair, and a looped out U. When the Rev peptide is bound to the RNA, the base pairs in the internal loop appear to be stabilized, although the RNA chemical shifts indicate that the RNA conformation undergoes some changes when bound by Rev peptide.
本文介绍了对一种30个核苷酸的RNA寡核苷酸（RBE3）的1H NMR研究，该寡核苷酸含有HIV-1 Rev反应元件（RRE）中Rev的高亲和力结合位点、RBE3的两种衍生物（RBE3AA和RBE3-A），以及RBE3与源自HIV-1 Rev RNA结合结构域的肽的复合物。RRE的高亲和力结合位点由一个不对称内环和周围的沃森-克里克碱基对组成。在野生型RRE中，其中一个茎被一个环封闭；在REB3中，这被稳定的UUCG四环所取代。NOE数据表明，游离RNA的内环包含基于体外筛选实验预测的结构特征[巴特尔，D.P.等人（1991年）《细胞》67卷，529 - 536页]。这些结构特征包括一个顺式G.反式G碱基对、一个反式G.反式A碱基对和一个环出的U。当Rev肽与RNA结合时，内环中的碱基对似乎得到了稳定，尽管RNA化学位移表明，当被Rev肽结合时，RNA构象会发生一些变化。
