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首页> 外文期刊>Journal of Molecular Biology >BULGE-INDUCED BENDS IN RNA - QUANTIFICATION BY TRANSIENT ELECTRIC BIREFRINGENCE
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BULGE-INDUCED BENDS IN RNA - QUANTIFICATION BY TRANSIENT ELECTRIC BIREFRINGENCE

机译：RNA凸出的弯曲-瞬态电双引定量

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摘要

Bulges represent one of the most common non-helical elements in RNA, often displaying a strong degree of phylogenetic conservation, both in location and sequence, within larger RNA molecules. Thus, knowledge of the conformation and flexibility of RNA bulges is an important prerequisite for understanding the rules governing the formation of tertiary structure within the larger molecules. In the current investigation, the magnitudes of the bends induced in a 148 base-pair duplex RNA molecule by single, centrally located bulges of varying size (n = 1 to 6) and base composition (A(n) and U-n series) have been determined through the use of transient electric birefringence (TEE). The TEE approach is highly sensitive to the changes in the global shape of RNA (or DNA) helices that accompany the introduction of points of bending or flexibility near the center of the helix. In the current instance, bulge angles deduced from TEE measurements ranged from similar to 7 degrees to similar to 93 degrees, with the angle increasing with increasing n for both A(n) and U-n series. For both A(n) and U-n series in the absence of Mg2+, the angle increment per added nucleotide varied from similar to 20 degrees to similar to 8 degrees as n increased from 1 to 6. These angle increments remained unchanged for the A(n) series in the presence of Mg2+; however, the angle increments for the U-n series were reduced by a factor of 2 for all values of n. Thus the current observations have identified structural transitions in one of the simplest non-helical elements in RNA, transitions that are dependent on both sequence and counterion valence. Finally, the measured bend angles are strongly correlated with the degree of reduction in electrophoretic mobility of bulge-containing RNA helices. The observed correlation was used to obtain a semi-empirical relationship between bend angle and mobility in order that additional angles might be assigned, by interpolation, through the use of gel data alone. [References: 56]
机译：凸起代表RNA中最常见的非螺旋元件之一，通常在较大的RNA分子中在位置和序列上都表现出高度的系统发育保守性。因此，了解RNA凸起的构象和柔韧性是理解控制较大分子内三级结构形成的规则的重要前提。在当前的研究中，由148个碱基对的双链RNA分子通过大小不同的单个凸起（n = 1至6）和碱基组成（A（n）和Un系列）诱导的弯曲幅度为通过使用瞬态双折射（TEE）来确定。 TEE方法对伴随螺旋中心附近弯曲点或柔韧性点引入的RNA（或DNA）螺旋整体形状的变化高度敏感。在当前情况下，从TEE测量得出的凸角范围从相似的7度到相似的93度，对于A（n）和U-n系列，该角度均随n的增加而增加。对于不存在Mg2 +的A（n）和Un系列，随着n从1增加到6，每个添加核苷酸的角度增量从相似的20度变化为相似的8度。这些角度增量对于A（n ）在Mg2 +存在下的系列;但是，对于所有n值，U-n系列的角度增量减少了2倍。因此，目前的观察已经鉴定出RNA中最简单的非螺旋元件之一中的结构转变，该转变既取决于序列价又取决于抗衡价。最后，所测得的弯曲角与含有凸起的RNA螺旋的电泳迁移率的降低程度密切相关。观察到的相关性用于获得弯曲角度和迁移率之间的半经验关系，以便可以通过插值仅通过使用凝胶数据来分配其他角度。 [参考：56]

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