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RNA TARGETING WITH TERTIARY INTERACTIONS

通过三级相互作用进行 RNA 靶向

基本信息

批准号：
6170348
负责人：
DOUGLAS H. TURNER
金额：
$ 7.98万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-01 至 2001-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6170348
关键词：
Pneumocystis carinii RNA binding protein RNA splicing antifungal agents antisense nucleic acid drug design /synthesis /production molecular site oligonucleotides phosphonate protein structure ribosomal RNA

项目摘要

DESCRIPTION: (Adapted from Applicant's Abstract) This RO3 application is in response to a Program Announcement entitled Innovative Drug Discovery Research in AIDS Opportunistic Infections. The ultimate goal of this project is to discover principles that allow targeting of RNA with small oligonucleotide based compounds that inhibit function. Establishment of such principles could have wide ranging applications, including design of compounds to treat HIV and opportunistic infections. New paradigms to be explored are (1) binding enhancement by tertiary interactions (BETI), (2) suicide inhibition, and (3) oligonucleotide-catalyzed hydrolysis of splice sites. The group I self-splicing intron from the ribosomal RNA precursor in Pneumocystis carinii will serve as the model system for these studies. Correct splicing of this precursor is required for formation of active ribosomes. Preliminary work has shown that a nuclease stable, N3'-P5' phosphoramidate hexanucleotide exhibits binding enhanced by tertiary interactions in binding tightly to the sequence that normally aligns the 5' splice site. This hexanucleotide inhibits self-splicing via a suicide inhibition mechanism. Specific aims for the proposed work are: (1) Determine if oligonucleotides with uncharged methylphosphonate linkages exhibit BETI and inhibition of self-splicing. (2) Determine if BETI is available at other sites on the group I intron. (3) Determine if in vitro oligonucleotide inhibitors are taken up by P. carinii cells. The results are expected to provide fundamental principles for designing inhibitors of many RNA functions. Because maturation of rRNA precursor is essential for proliferation of P. carinii, and no group I self-splicing introns are known in human genes, the results could also provide potential therapeutics for P. carinii pneumonia.

描述：（改编自申请人的摘要）此 RO3 申请位于对题为“创新药物发现研究”的计划公告的回应艾滋病机会性感染。该项目的最终目标是发现用小寡核苷酸靶向 RNA 的原理抑制功能的化合物。制定此类原则可以具有广泛的应用，包括设计治疗艾滋病毒和机会性感染。需要探索的新范式是（1）绑定通过三级相互作用增强（BETI），（2）自杀抑制，以及（3）寡核苷酸催化的剪接位点水解。 I组自剪接卡氏肺囊虫核糖体 RNA 前体的内含子将作为这些研究的模型系统。该前体的正确剪接是形成活性核糖体所需的。初步工作表明，核酸酶稳定，N3'-P5' 氨基磷酸酯六核苷酸表现出结合通过与序列紧密结合的三级相互作用增强通常对齐 5' 剪接位点。这种六核苷酸抑制自我剪接通过自杀抑制机制。拟议工作的具体目标是： (1) 确定寡核苷酸是否具有不带电荷的甲基膦酸酯键表现出 BETI 和自剪接抑制。 (2) 判断BETI是否为可在 I 组内含子上的其他位点获得。 (3) 判断是否在体外寡核苷酸抑制剂被卡氏疟原虫细胞吸收。结果是有望为设计许多 RNA 抑制剂提供基本原理功能。因为 rRNA 前体的成熟对于增殖至关重要 P. carinii，并且在人类基因中没有已知的 I 组自剪接内含子，该结果还可以为卡氏疟原虫肺炎提供潜在的治疗方法。