Hoechst 33258 selectively inhibits group I intron self-splicing by affecting RNA folding
Hoechst 33258 selectively inhibits group I intron self-splicing by affecting RNA folding
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DOI：
10.1002/cbic.200400159
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发表时间：
2004-12-03
期刊：
影响因子：
3.2
通讯作者：
Turner, DH
中科院分区：
文献类型：
--
作者：
Disney, MD;Childs, JL;Turner, DH
由于耐药菌株的增加和免疫功能低下的人的数量，真菌病原体的流行正在增加。白念珠菌是其中一种致病菌，约40%的菌株在其大亚基rRNA前体中含有I组自我剪接内含子，这是一个潜在的RNA药物靶点。在此，我们报道Hoechst 33258及其衍生物是C.白色念珠菌I组内含子自剪接，在2 mM Mg 2+中的IC 50为17 μ M。在Hoechst 33258存在下的内含子的化学探测揭示了内含子的P4/P6区域中的几个核苷酸的折叠受到影响。在Hoechst 33258的存在下，J 4/5区域附近的核苷酸被保护免于化学修饰，并且附近的几个核苷酸更具反应性;这表明该区域是分子的结合位点。这些结果扩展了关于RNA的小分子靶向的可用信息，并表明Hoechst提供的RNA靶向支架可能在设计抑制RNA功能的化合物方面证明是有价值的。
Fungal pathogens are increasing in prevalence due to an increase in resistant strains and the number of immunocompromised humans. Candida albicans is one of these pathogens, and similar to 40 % of strains contain a group I self-splicing intron, which is a potential RNA drug target, in their large subunit rRNA precursor. Here, we report that Hoechst 33258 and derivatives thereof are selective inhibitors of C. albicans group I intron self-splicing with on IC50 of 17 muM in 2 mM Mg2+. Chemical probing of the intron in the presence of Hoechst 33258 reveals that the folding of several nucleotides in the P4/P6 region of the intron is affected. A nucleotide near the J4/5 region is protected from chemical modification in the presence of Hoechst 33258 and several nearby are more reactive; this suggests that this region is the molecule's binding site. These results expand the available information on small-molecule targeting of RNA and suggest that the RNA-targeting scaffold provided by Hoechst may prove valuable in designing compounds that inhibit the functions of RNA.