Rational design of ligands targeting triplet repeating transcripts that cause RNA dominant disease: application to myotonic muscular dystrophy type 1 and spinocerebellar ataxia type 3.
Rational design of ligands targeting triplet repeating transcripts that cause RNA dominant disease: application to myotonic muscular dystrophy type 1 and spinocerebellar ataxia type 3.
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DOI：
10.1021/ja9020149
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发表时间：
2009-07-22
影响因子：
15
通讯作者：
Disney MD
中科院分区：
文献类型：
--
作者：
Pushechnikov A;Lee MM;Childs-Disney JL;Sobczak K;French JM;Thornton CA;Disney MD
在此，我们描述了高亲和力配体的设计，结合扩展的rCUG和rCAG重复RNA表达的强直性肌营养不良和脊髓小脑共济失调。这些配体还以纳摩尔IC 50抑制与两种疾病有关的RNA-蛋白质复合物的形成。扩增的rCUG和rCAG重复序列形成稳定的RNA发夹，在茎中有规律地重复内环，并对细胞功能产生有害影响。结合重复序列的配体显示双苯并咪唑Hoechst 33258的衍生物，其通过搜索已知的RNA-配体相互作用来鉴定。合成了一系列13个模块化组装的配体，配体模块之间具有定义的化合价和距离，以同时靶向这些RNA中的多个基序。最亲合力的结合剂五聚体以13 nM的Kd结合rCUG-重复发夹。与一系列相关RNA相比，五聚体以4.4至>200倍的特异性结合rCUG重复序列。此外，与rCUG重复序列结合的亲和力随着化合价的增加而增加，而与基因组DNA结合的背景相应减少。然后，确定多价配体是否抑制肌盲样1（MBNL 1）蛋白对RNA重复的识别，所述MBNL 1蛋白是其螯合导致DM 1中的剪接缺陷的扩增的rCUG结合蛋白。在具有纳摩尔IC 50的几种化合物中，最有效的抑制剂是五聚体，其也抑制rCAG重复序列-MBNL 1复合物的形成。设计的合成配体和MBNL 1与重复RNA的结合数据的比较表明，合成配体对DM 1 RNA的亲和力比MBNL 1高23倍，并且对DM 1 RNA的特异性更高。进一步的研究表明，所设计的配体对小鼠成肌细胞具有细胞渗透性。因此，已经设计了结合重复RNA的细胞可渗透配体，其表现出比天然蛋白质更高的结合RNA的亲和力和特异性。这些研究提出了一种靶向RNA的通用方法，包括那些导致RNA显性疾病的RNA。
Herein, we describe the design of high affinity ligands that bind expanded rCUG- and rCAG-repeat RNAs expressed in myotonic dystrophy and spinocerebellar ataxia. These ligands also inhibit, with nanomolar IC50's, the formation of RNA-protein complexes that are implicated in both disorders. The expanded rCUG and rCAG repeats form stable RNA hairpins with regularly repeating internal loops in the stem and have deleterious effects on cell function. The ligands that bind the repeats display a derivative of the bis-benzimidazole Hoechst 33258, which was identified by searching known RNA-ligand interactions. A series of 13 modularly assembled ligands with defined valencies and distances between ligand modules was synthesized to target multiple motifs in these RNAs simultaneously. The most avid binder, a pentamer, binds the rCUG-repeat hairpin with a Kd of 13 nM. As compared to a series of related RNAs, the pentamer binds to rCUG-repeats with 4.4- to >200-fold specificity. Furthermore, the affinity of binding to rCUG-repeats shows incremental gains with increasing valency while the background binding to genomic DNA is correspondingly reduced. Then, it was determined whether the multivalent ligands inhibit the recognition of RNA repeats by Muscleblind-like 1 (MBNL1) protein, the expanded-rCUG binding protein whose sequestration leads to splicing defects in DM1. Among several compounds with nanomolar IC50's, the most potent inhibitor is the pentamer, which also inhibits the formation of rCAG repeat-MBNL1 complexes. Comparison of the binding data of the designed synthetic ligands and MBNL1 to repeating RNAs shows that the synthetic ligand is 23-fold higher affinity and more specific to DM1 RNAs than MBNL1. Further studies show that the designed ligands are cell permeable to mouse myoblasts. Thus, cell permeable ligands that bind repetitive RNAs have been designed that exhibit higher affinity and specificity for binding RNA than natural proteins. These studies suggest a general approach to targeting RNA, including those that cause RNA dominant disease.
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能够在体外抑制（CUG）重复RNA-MBNL1相互作用的分子的动态组合选择：发现靶向肌发育症的铅化合物（DM1）。
影响因子：
15
作者：
Gareiss, Peter C.;Sobczak, Krzysztof;McNaughton, Brian R.;Palde, Prakash B.;Thornton, Charles A.;Miller, Benjamin L.
通讯作者：
Miller, Benjamin L.
影响因子：
2.9
作者：
Aminova, Olga;Paul, Dustin J.;Disney, Matthew D.
通讯作者：
Disney, Matthew D.
DOI：
10.1073/pnas.2035074100
发表时间：
2003-10-14
影响因子：
11.1
作者：
Best, TP;Edelson, BS;Dervan, PB
通讯作者：
Dervan, PB
影响因子：
3.5
作者：
Belitsky, JM;Leslie, SJ;Dervan, PB
通讯作者：
Dervan, PB
DOI：
10.1073/pnas.97.17.9367
发表时间：
2000-08-15
影响因子：
11.1
作者：
Erlanson, DA;Braisted, AC;Wells, JA
通讯作者：
Wells, JA