Interaction of Aminoglycosides with Human Mitochondrial 12S rRNA Carrying the Deafness-Associated Mutation
Interaction of Aminoglycosides with Human Mitochondrial 12S rRNA Carrying the Deafness-Associated Mutation
复制标题
DOI：
10.1128/aac.00965-08
复制
发表时间：
2009-11-01
影响因子：
4.9
通讯作者：
Guan, Min-Xin
中科院分区：
文献类型：
--
作者：
Qian, Yaping;Guan, Min-Xin
线粒体12S rRNA A1555G突变是氨基糖苷类药物致非综合征性耳聋的重要原因之一。在这里，我们使用了一种RNA引导的化学修饰方法来了解氨基糖苷类药物所致听力损失的发病机制。硫酸二甲酯(DMS)对携带A1555G突变的RNA寡核苷酸的化学修饰模式与在氨基糖苷类化合物存在下携带野生型序列的RNA寡核苷酸不同。在携带A1555G突变的RNA类似物中，碱基G1555以及碱基C1556和A1553在帕罗霉素、新霉素、庆大霉素、卡那霉素、妥布霉素或链霉素存在下对DMS的反应性降低。特别是，碱基G1555在0.1亩M到100亩M新霉素、庆大霉素或卡那霉素存在时表现出显著但相似的保护水平。相反，碱基G1555的保护水平似乎与帕罗霉素、妥布霉素或步进霉素的浓度有关。此外，携带A1555G突变的RNA类似物中的碱基A1492、C1493、C1494和A1557在这些氨基糖苷类化合物存在的情况下对DMS的反应性增加。这些数据表明，A1555G突变改变了12S rRNA A位氨基糖苷类化合物的结合特性，导致携带A1555G突变的12S rRNA局部构象发生变化。氨基糖苷类药物与携带A1555G突变的12S rRNA之间的相互作用为研究氨基糖苷类药物耳毒性的发病机制提供了新的思路。
The mitochondrial 12S rRNA A1555G mutation is one of the important causes of aminoglycoside-induced and nonsyndromic hearing loss. Here we employed an RNA-directed chemical-modification approach to understanding the pathogenesis of aminoglycoside-induced hearing loss. The patterns of chemical modification of RNA oligonucleotides carrying the A1555G mutation by dimethyl sulfate (DMS) were distinct from those of the RNA oligonucleotides carrying wild-type sequence in the presence of aminoglycosides. In the RNA analogue carrying the A1555G mutation, reduced reactivity to DMS occurred in base G1555 as well as in bases C1556 and A1553 in the presence of paromomycin, neomycin, gentamicin, kanamycin, tobramycin, or streptomycin. In particular, base G1555 exhibited marked but similar levels of protection in the presence of 0.1 mu M to 100 mu M neomycin, gentamicin, or kanamycin. In contrast, the levels of protection in base G1555 appeared to be correlated with the concentration of paromycin, tobramycin, or steptomycin. Furthermore, increasing reactivities to DMS in the presence of these aminoglycosides were observed for bases A1492, C1493, C1494, and A1557 in the RNA analogue carrying the A1555G mutation. These data suggested that the A1555G mutation altered the binding properties of aminoglycosides at the A site of 12S rRNA and led to local conformational changes in 12S rRNA carrying the A1555G mutation. The interaction between aminoglycosides and 12S rRNA carrying the A1555G mutation provides new insight into the pathogenesis of aminoglycoside ototoxicity.