Kinetoplastid SL RNA biogenesis

动质体 SL RNA 生物发生

基本信息

批准号：
7196546
负责人：
DAVID A CAMPBELL
金额：
$ 35.34万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7196546
关键词：
Address Affect African Trypanosomiasis Agriculture Base Pairing Binding Biogenesis Bioinformatics Candidate Disease Gene Cell Nucleus Cells Chagas Disease Complement Complex Data Elements Excision Exons Exonuclease Family Future Gene Expression Genes Genetic Translation Genome Goals Human In Vitro Insect Vectors Intervention Kinetoplastida Laboratory Organism Lead Leishmania Leishmaniasis Maps Medical Messenger RNA Methylation Methyltransferase Modeling Modification Molecular Mutation Nuclear Nuclear Export Organism Parasites Pathway interactions Pharmaceutical Preparations Phase Phenotype Play Poly U Positioning Attribute Principal Investigator Process Proteins Publishing RNA RNA Interference RNA Processing RNA Sequences RNA Splicing Reaction Research Ribose Role Series Small RNA Spliced Leader RNA Spliced Leader Sequences Structure Trans-Splicing Translations Trypanosoma brucei brucei Trypanosomatina Work base exportin 1 protein knock-down mRNA Stability nuclease pathogen programs research study spleen exonuclease stem therapeutic target trafficking

项目摘要

DESCRIPTION (provided by the applicant): Our research is aimed at understanding gene expression in the family Trypanosomatidae, which includes the parasitic protozoon's responsible for leishmaniasis, African Sleeping Sickness, and Chagas Disease. The focus is on the genesis and function of the spliced leader (SL) RNA, a small RNA that contributes the 5'-end sequence to every nuclear messenger RNA via a trans-splicing reaction. Trans-splicing, which is necessary for the conversion of polycistronic pre-messenger RNA (a likely consequence of the unusual genome organization found in kinetoplastids) into monocistronic mRNA, is not found in the human host or insect vector and thus represents a possible therapeutic target. This proposal outlines three complementary sets of experiments that detail the maturation pathway of the SL RNA common to Leishmania tarentolae and Trypanosoma brucei. 1) To refine the map of cis elements (SL RNA sequences and structures) necessary for accurate and efficient maturation and trans-splicing. Data obtained will allow the future identification of trans factors (protein and RNA) that interact with essential regions of RNA. 2) To isolate and examine the role of an XPO1-dependent nuclear export complex in SL RNA trafficking. 3) Experimental examination of the roles of a 3'-exonuclease and two ribose 2'-O- methyltransferases in the maturation of the 3' and 5' ends of the SL RNA, and bioinformatic identification and modeling of these proteins' interaction with the SL RNA. These three goals address discrete steps in the maturation of the SL RNA that determine its subsequent function. Specific inhibition of the trans-splicing pathway is lethal to the cell and thus a prime target for anti-parasite intervention.

描述（由申请人提供）：我们的研究旨在理解家族锥虫科中的基因表达，其中包括寄生虫原生动物负责利什曼病，非洲睡眠疾病和chagas病。重点是剪接领导者（SL）RNA的起源和功能，这是一种小的RNA，通过移植反应为每个核使者RNA贡献了5'-end序列。在人宿主或昆虫载体中未发现，这对于多用途预选前RNA的转化（可能是在动质塑料中发现的异常基因组组织的可能结果）的跨性变速器（可能是在动质塑料中发现的不寻常的结果），因此代表了可能的疗法靶标。该提议概述了三组互补的实验集，详细介绍了Leishmania tarentolae和Brucei的SL RNA的成熟途径。 1）为准确有效的成熟和跨拼接所必需的顺式元素（SL RNA序列和结构）的图。获得的数据将允许将来鉴定与RNA基本区域相互作用的反式因子（蛋白质和RNA）。 2）分离并检查XPO1依赖性核出口复合物在SL RNA运输中的作用。 3）对SL RNA的3'和5'端的成熟中的3'-核酸酶和两个核糖2'-甲基转移酶的作用的实验检查，以及这些蛋白质与SL RNA的相互作用的生物信息学鉴定和建模。这三个目标介绍了确定其后续功能的SL RNA成熟的离散步骤。对移植途径的特异性抑制对细胞是致命的，因此是抗寄生虫干预的主要靶标。