Costabilization of peptide and RNA structure in an HIV Rev peptide-RRE complex.
Costabilization of peptide and RNA structure in an HIV Rev peptide-RRE complex.
复制标题
HIV Rev 肽-RRE 复合物中肽和 RNA 结构的共稳定。
DOI：
10.1021/bi00252a025
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Frankel,AD
中科院分区：
文献类型：
--
作者：
Tan,R;Frankel,AD
1994年9月30日接收的修订版Mandarin摘要：一种富含精氨酸的肽，对应于人类免疫缺陷病毒Rev蛋白的氨基酸34-50，当该肽处于α-螺旋构象时，已显示出特异性结合其RNA结合位点（RRE）。六个氨基酸（Thr 34、Arg 35、Arg 38、Arg 39、Asn 40或Arg 44）中的任何一个的突变显示出在体外强烈降低特异性RNA结合亲和力，这表明这些残基可能接触RNA的特异性碱基或独特的结构特征。我们现在表明，四个精氨酸侧链，而不仅仅是他们的电荷，是重要的特异性结合在体内，并提出证据表明，三个额外的精氨酸（Arg 46，Arg 48和Arg 50）maymake静电接触的RRE。Rev肽的RNA结合特异性在体外具有温度依赖性，与α-螺旋解折叠相关。圆二色谱实验表明，肽的螺旋结构是稳定的，当结合特异性的RRE和RNA的结合后经历构象变化。由于在这个模型系统中肽和RNA的结构在结合时是相互稳定的，因此建议整个复合物可以被看作是一个单一的折叠单元。从生物学的角度来看，RNA-蛋白质识别的研究是有趣的，因为许多细胞功能，包括转录，RNA剪接和翻译，都依赖于蛋白质和RNA的特异性相互作用。从大分子的角度来看，这个问题很有趣，因为RNA可以折叠成各种各样的三级结构，尽管到目前为止只有少数结构已经解决。tRNA的三维结构已经被人们所知有相当长的一段时间了，现在已经知道了三种tRNA合成酶-tRNA复合物的共晶结构
Revised Manuscript Received September 30, 1994® abstract: An arginine-rich peptide corresponding to amino acids 34-50 of the human immunodeficiency virus Rev protein has beenshown to bind specifically to its RNA-binding site (RRE) when the peptide is in an a-helical conformation. Mutation of any one of six amino acids (Thr34, Arg35, Arg38, Arg39, Asn40, or Arg44) was shown to strongly decrease specific RNA-binding affinity in vitro, suggesting that these residues may contact specific bases or distinct structural features of the RNA. We now show that the four arginine side chains, and not just their charge, are important for specific binding in vivo, and present evidence that three additional arginines (Arg46, Arg48, and Arg50) maymake electrostatic contacts to the RRE. RNA-binding specificity of the Rev peptide is temperature-dependent in vitro, correlating with a-helix unfolding. Circular dichroismexperiments indicate that the peptide helical structure is stabilized when bound specifically to the RRE and that the RNA undergoes a conformational change upon binding. Because the structures of the peptide and RNA in this model systemappear to be mutually stabilized upon binding, it is suggested that the entire complex may be viewed as a single folding unit.From a biological perspective, the study of RNA—protein recognition is interesting because many cellular functions, including transcription, RNA splicing, and translation, depend on the specific interaction of proteins and RNA. From a macromolecular perspective, the problem is interesting because RNAs can fold into a wide range of tertiary structures, although so far only a few structures have been solved. The three-dimensional structure of tRNA has been known for quite some time, and the cocrystal structures of three tRNA synthetase—tRNA complexes are now known