Biochemical characterization of the kink-turn RNA motif
Biochemical characterization of the kink-turn RNA motif
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DOI：
10.1093/nar/gkg760
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发表时间：
2003-10-01
影响因子：
14.9
通讯作者：
Inoue, T
中科院分区：
文献类型：
--
作者：
Matsumura, S;Ikawa, Y;Inoue, T
RNA作为传递遗传信息的媒介，在细胞中发挥着多种作用。与蛋白质一样，阐明RNA的三维（3D）结构对于理解它们的各种作用是重要的。结晶核糖体的原子结构的测定使以前未知的RNA结构基序的鉴定成为可能。在RNA双螺旋中引起急剧弯曲的扭结-转角（K-turn或GA）基序被鉴定为这些结构基序之一。为了生物化学表征K-转角，将基序插入P4-P6 RNA的铰链区，这是最广泛研究的自折叠RNA，并研究其性质。使用非变性PAGE和硫酸二甲酯（DMS）修饰分析含有三种不同K-转角基序的构建体的稳定性和金属离子需求。尖锐弯曲结构的形成取决于二价阳离子如Mg 2+或Ca 2+的存在，尽管其所需浓度对于每个基序是不同的。
RNA, which acts as a medium for transmitting genetic information, plays a variety of roles in a cell. As with proteins, elucidation of the three- dimensional (3D) structures of RNAs is important for understanding their various roles. Determination of the atomic structures of crystallized ribosome has enabled the identification of previously unknown RNA structural motifs. The kink-turn (K-turn or GA) motif, which causes a sharp bend in an RNA double helix, was identified as one of these structural motifs. To biochemically characterize the K-turn, the motif was inserted into a hinge region of P4-P6 RNA, which is the most extensively studied self-folding RNA, and its properties were investigated. The stability and metal ion requirement of the constructs containing three different K-turn motifs were analyzed using native PAGE and dimethyl sulfate (DMS) modification. The formation of the sharp bending structure depends on the presence of divalent cation like Mg2+ or Ca2+, although its required concentration is different for each motif.