Ribonucleopeptides: Functional RNA-peptide complexes
Ribonucleopeptides: Functional RNA-peptide complexes
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DOI：
10.1002/bip.10576
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发表时间：
2004-01-01
期刊：
影响因子：
2.9
通讯作者：
Morii, T
中科院分区：
文献类型：
--
作者：
Hagihara, M;Hasegawa, T;Morii, T
从随机化的RNA分子库中选择功能性RNA成功地提供了特异性结合小分子并具有催化活性的RNA适体。最近的核糖体RNA复合物的结构分析表明，RNA-蛋白质复合物将是一个新的结构候选人的设计定制的受体和酶。我们通过基于结构的设计方法和体外筛选方法设计了一个由RNA亚基和肽亚基组成的ATP结合结构域。RNA亚基被设计成由两个功能结构域组成;具有20个随机核苷酸的ATP结合结构域和作为RNA结合肽的结合位点的相邻的stein区域。将随机化核苷酸区域置于HIV-1 Rev反应元件旁边，以在Rev肽存在下形成“核糖核肽”池。从随机化池中体外选择RNA寡核苷酸提供了对ATP特异性的核糖核肽受体。ATP结合核糖核酸肽不共享ATP适体的已知共有核苷酸序列，并且在不存在Rev肽的情况下完全失去其ATP结合能力。的核糖核酸肽的ATP结合活性增加的N-末端氨基酸的Rev肽的取代。这些结果表明，该肽稳定了RNA的功能结构，并表明该肽的RNA结合区外的氨基酸参与ATP结合。我们的方法将提供一个新的策略，为定制的核糖核肽受体的设计。(C)2004 Wiley Periodicals，Inc.
Selection of functional RNAs from randomized pool of RNA molecules successfully affords RNA aptamers that specifically bind to small molecules, and that have catalytic activities. Recent structural analyses of the ribosomal RNA complex suggest that the RNA-protein complex would be a new structural candidate for the design of tailor-made receptors and enzymes. We have designed an ATP binding domain that consists of an RNA subunit and a peptide subunit by means of structure-based design approach and successive in vitro selection method. The RNA subunit is designed to consist of two functional domains; an ATP binding domain with 20 randomized nucleotides and an adjacent stein region that serves as a binding site for the RNA-binding peptide. The randomized nucleotide region was placed next to the HIV-1 Rev response element to enable the formation of "ribonucleopeptide" pools in the presence of the Rev peptide. In vitro selection of RNA oligonucleotides from the randomized pool afforded a ribonucleopeptide receptor specific for ATP. The ATP-binding ribonucleopeptide did not share the known consensus nucleotide sequence for ATP aptamers, and completely lost its ATP-binding ability in the absence of the Rev peptide. The ATP-binding activity of the ribonucleopeptide was increased by a substitution of the N-terminal amino acid of the Rev peptide. These results demonstrate that the peptide stabilizes the functional structure of RNA and suggest that amino acids outside the RNA binding region of the peptide participate in the ATP binding. Our approach would provide a new strategy for the design of tailor-made ribonucleopeptide receptors. (C) 2004 Wiley Periodicals, Inc.