SPECIFIC BINDING OF AMINOGLYCOSIDE ANTIBIOTICS TO RNA
SPECIFIC BINDING OF AMINOGLYCOSIDE ANTIBIOTICS TO RNA
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DOI：
10.1016/1074-5521(95)90047-0
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发表时间：
1995-05-01
影响因子：
--
通讯作者：
RANDO, RR
中科院分区：
文献类型：
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作者：
WANG, Y;RANDO, RR
背景：氨基糖苷类抗生素干扰核糖体蛋白合成和内含子剪接。各种证据表明RNA是氨基糖苷类药物的分子靶点，但对识别过程知之甚少。特定氨基糖苷类的识别是否对特定RNA结构具有特异性？如果是，承认的规则是什么？我们已经开始研究这个问题，通过在体外选择的RNA分子，可以特异性地结合到氨基糖苷类抗生素妥布霉素。结果：一个RNA多样性库被用来选择能够结合到氨基糖苷类抗生素妥布霉素的序列。经过6个循环的选择，82%的RNA特异性结合妥布霉素。将选择的RNA逆转录成DNA，然后克隆。在低选择严格性下，数量级为10（7）的极大量克隆产生能够结合妥布霉素的RNA，K（d）在μ M范围内（与妥布霉素与大肠杆菌核糖体结合时观察到的值相似）。18个克隆的测序显示没有明显的共有序列。在较高的选择stringencies（K（d）的nM范围内），只有两个共识序列的bindings.Conclusions：我们已经表明，RNA分子可以很容易地选择结合氨基糖苷类妥布霉素。以高亲和力结合妥布霉素的RNA含有可限于预测的茎环结构的共有结合区。这些研究为理解RNA-氨基糖苷类识别的基础开辟了道路。
Background: Aminoglycoside antibiotics interfere with ribosomal protein synthesis and with intron splicing. Various lines of evidence suggest that RNA is the molecular target for aminoglycosides, but little is known about the recognition process. Is recognition of a particular aminoglycoside specific for certain RNA structures? If so, what are the rules for recognition? We have begun to investigate this problem by in vitro selection of RNA molecules that can specifically bind to the aminoglycoside antibiotic tobramycin.Results: An RNA diversity library was used to select for sequences capable of binding to the aminoglycoside antibiotic tobramycin. After six cycles of selection, 82 % of the RNA bound to tobramycin specifically. The selected RNA was reverse-transcribed into DNA, which was then cloned. At low selection stringency, an extremely large number of clones, on the order of 10(7), produced RNAs capable of binding tobramycin with K(d)s in the mu M range (values similar to that observed for the binding of tobramycin to Escherichia coli ribosomes). Sequencing of 18 of the clones revealed no obvious consensus sequence. At higher selection stringencies (K(d)s in the nM range) only two consensus sequences for binding were observed.Conclusions: We have shown that RNA molecules can be readily selected that bind the aminoglycoside tobramycin. The RNAs that bind tobramycin with high affinity contain consensus binding regions that may be confined to predicted stern-loop structures. These studies open the way for understanding the basis of RNA-aminoglycoside recognition.