Design, assembly, and evaluation of RNA-protein nanostructures.
Design, assembly, and evaluation of RNA-protein nanostructures.
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DOI：
10.1007/978-1-4939-2562-9_14
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发表时间：
2015
影响因子：
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通讯作者：
Hirohisa Ohno;E. Osada;Hirohide Saito
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文献类型：
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作者：
Hirohisa Ohno;E. Osada;Hirohide Saito
利用RNA -蛋白相互作用基序（RNP基序）来设计和构建纳米级物体具有扩展RNA纳米技术领域的潜力。原则上，RNP基序可以很容易地整合到RNA纳米物体中，为增加RNA的功能和结构复杂性提供了一种替代技术。研究RNP纳米结构的设计原理，将有助于从零开始构建高度复杂的生物大分子复合物，如核糖体。作为实现这一目标的第一步，我们通过使用盒C/D扭结旋转(K-turn)-L7Ae RNP基序设计和构建了三角形纳米结构。我们发现K-turn RNA和核糖体蛋白L7Ae可以形成一个形状像等边三角形的纳米结构，由附着在RNA支架尖端的三种蛋白质组成。该复合物的构建依赖于L7Ae与RNA中K-turn基序的结合。RNP基序允许RNA在三个位置弯曲约60°，形成纳米级三角形。功能RNP三角形在三个尖端处具有所需的蛋白质模块，可以以模块化的方式构建。在这里，我们描述了如何设计、构建和评估RNP纳米结构。
The use of RNA–protein interaction motifs (RNP motifs) to design and build nanoscale objects has the potential to expand the field of RNA nanotechnology. In principle, RNP motifs can be integrated easily into RNA nano objects, providing an alternative technique to increase the functional and structural complexities of the RNA. Investigating the design principles of RNP nanostructures will enable the construction of highly sophisticated biomacromolecular complexes such as ribosomes from scratch. As an initial step towards this goal, we designed and constructed triangular-like nanostructures by employing box C/D kink-turn (K-turn)-L7Ae RNP motifs. We showed that the K-turn RNA and the ribosomal protein L7Ae could form a nanostructure shaped like an equilateral triangle that consists of the three proteins attached to the tips of the RNA scaffold. The construction of the complex depends on L7Ae binding to the K-turn motifs in the RNA. The RNP motif allows the RNA to bend by approximately 60° at three positions to form a nanoscale triangle. Functional RNP triangles with desired protein modules at the three tips can be constructed in a modular manner. Here, we describe how to design, construct, and evaluate the RNP nanostructures.