The translation apparatus of Leishmania: from basic analysis to pursuit of novel

利什曼原虫的翻译机构：从基础分析到小说追求

基本信息

批准号：
9297201
负责人：
GERHARD WAGNER
金额：
$ 34.87万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-06 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9297201
关键词：
Adverse effects Affect Appearance Back Binding Biochemical Biological Assay Biology Bite Chemicals Chemistry Clinical Code Complex Country Crystallization Data Development Disease Drug Targeting Effectiveness Environment Epitopes Eukaryota Family Female Fluorescence Fluorescence Polarization Genes Genome Geography Goals Human In Vitro Infection Institutes Interruption Intravenous Leishmania Leishmania major Leishmaniasis Length Life Ligands Light Malignant Neoplasms Mammalian Cell Measurement Measures Messenger RNA Molecular Molecular Biology Monitor NMR Spectroscopy Organism Outcome Parasite resistance Parasites Parasitology Peptide Initiation Factors Peptides Pharmacotherapy Phlebotominae Plasmodium falciparum Protein Biosynthesis Proteins Proteolysis RNA Cap-Binding Proteins Recruitment Activity Relaxation Research Research Proposals Residual state Resistance Resolution Ribosomes Roentgen Rays Role Route Scaffolding Protein Sequence Homology Site Stress Structure Syndrome System Time Translation Initiation Translations Trypanosoma brucei brucei Trypanosoma cruzi Trypanosomatina X-Ray Crystallography base biophysical techniques effective therapy experimental study flexibility high throughput screening inhibitor/antagonist medical attention medical schools member neglect new therapeutic target novel novel therapeutics pathogen protein complex protein protein interaction public health relevance screening small molecule inhibitor tool

项目摘要

DESCRIPTION (provided by applicant): Protozoan parasites that belong to the genus Leishmania from the Trypanosomadae family, causes Leishmaniasis in humans, which affects 350 million people in 88 countries worldwide. The parasites are principally transmitted to humans following the bite of female phlebotomine sandflies, which carry the infectious stage of the organism. Leishmaniasis comprise three major clinical syndromes principally determined by the species of infecting parasites, which vary geographically. Few therapies are currently available, but strong side effects and parasite resistance limit their effectiveness. The recent advancements in determining several trypanosomatid genomes provided an invaluable tool to gather information regarding the molecular and biochemical mechanisms used by these organisms. However, it has proven difficult to correctly assign specific functions to gene coding sequences because the sequence homology between these parasites and their higher eukaryotic counterparts is extremely low. Until recently, little effort was invested in the Leishmania cap-binding complexes in the human stage, which are required to initiate protein synthesis in parasites. In this proposal, we focus on three translation initiation factors from Leishmania major in the human stage: LeishIF4E-1, Leish4E-IP and LeishIF3. While LeishIF4E-1 has emerged as the crucial cap-binding protein, it is hypothesized that Leish4E-IP and LeishIF3 are, respectively, a regulator of translation initiation and a scaffold protein responsibl for recruiting the small ribosomal subunit at the 5' end of messenger RNAs. To investigate whether this is the case, we will apply a combination of biophysical techniques, including NMR spectroscopy and X-ray crystallography, and molecular biology approaches to study the structure/function of these factors in vitro and within parasites. Once we determine indispensable interactions that promote translation initiation in the parasite, we will explore these potential targets in high-throughput (HTP) small molecule inhibitor screens. This proposal will allow us to highlight similarities and differences between translation initiation in human cels and Leishmania parasites, and validate the Leishmania translation machinery as a potential drug target. This concept is backed by data obtained previously in the Wagner lab where a crucial interaction between two human translation initiation factors (eIF4E/eIF4G) has been targeted in a HTP screen and led to the discovery of compounds with anti-cancer activity. The research will pursue three specific aims: 1. Determine the structure of the LeishIF4E-1 complex with its ligand cap-4 and Leish4E-IP. 2. Characterize the LeishIF3 recruitment to the Leishmania cap-binding complex. 3. Develop assays for HTP screens in search of Leishmania inhibitory compounds.

描述（由申请人提供）：属于锥虫家族的利什曼尼亚属的原生动物寄生虫会导致人类利什曼病，这影响了全球88个国家的3.5亿人。这些寄生虫主要是在雌性静脉菌粒沙叶的咬伤后传播给人类的，这些沙蝇带有有机体的传染性阶段。利什曼病包括三种主要的临床综合征，主要由感染寄生虫的物种确定，它们在地理上变化。目前很少有疗法，但是强烈的副作用和寄生虫的抗性限制了其有效性。确定几种锥形基因组的最新进展为收集有关这些生物使用的分子和生化机制的信息提供了宝贵的工具。但是，事实证明，很难将特定功能正确分配给基因编码序列，因为这些寄生虫与它们较高的真核生物对应物之间的序列同源性极低。直到最近，在人类阶段的利什曼尼亚帽结合综合体中投入了很少的努力，这是在寄生虫中启动蛋白质合成所必需的。在此提案中，我们将重点放在人类阶段利什曼尼亚专业的三个翻译起始因素上：Leishif4e-1，Leish4e-IP和Leishif3。虽然Leishif4e-1已成为关键的盖结结蛋白，但假设Leish4e-IP和Leishif3分别是翻译起始的调节剂和脚手架蛋白质的责任，用于招募Messenger RNA的5'末端的小核糖体亚基。为了研究这种情况，我们将采用生物物理技术的组合，包括NMR光谱和X射线晶体学，以及分子生物学方法来研究这些因素在体外和寄生虫中的结构/功能。一旦我们确定了促进寄生虫翻译起始的必不可少的相互作用，我们将在高通量（HTP）小分子抑制剂筛选中探索这些潜在靶标。该提案将使我们能够强调人类卡尔斯和利什曼原虫寄生虫的翻译启动之间的相似性和差异，并验证利什曼原虫翻译机械作为潜在的药物靶标。这个概念得到了先前在Wagner Lab中获得的数据支持，在Wagner Lab中，在HTP筛选中已针对两个人类翻译起始因子（EIF4E/EIF4G）之间的关键相互作用，并导致发现具有抗癌活性的化合物。该研究将追求三个具体目标：1。确定Leishif4e-1复合物与配体CAP-4和LEISH4E-IP的结构。 2。将Leishif3招募到Leishmania Cap结合综合体中。 3。开发HTP筛选的测定法，以寻找利什曼尼亚抑制性化合物。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structural basis for LeishIF4E-1 modulation by an interacting protein in the human parasite Leishmania major.

DOI：
10.1093/nar/gky194
发表时间：
2018-04-20
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Meleppattu S;Arthanari H;Zinoviev A;Boeszoermenyi A;Wagner G;Shapira M;Léger-Abraham M
通讯作者：
Léger-Abraham M

The eIF3 complex of Leishmania-subunit composition and mode of recruitment to different cap-binding complexes.

利什曼原虫亚基组成的 eIF3 复合物以及招募不同帽结合复合物的模式。

DOI：
10.1093/nar/gkv564
发表时间：
2015
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Meleppattu,Shimi;Kamus-Elimeleh,Dikla;Zinoviev,Alexandra;Cohen-Mor,Shahar;Orr,Irit;Shapira,Michal
通讯作者：
Shapira,Michal

A newly identified Leishmania IF4E-interacting protein, Leish4E-IP2, modulates the activity of cap-binding protein paralogs.

新鉴定的利什曼原虫 IF4E 相互作用蛋白 Leish4E-IP2 可调节帽结合蛋白旁系同源物的活性。

DOI：
10.1093/nar/gkaa173
发表时间：
2020
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Tupperwar,Nitin;Meleppattu,Shimi;Shrivastava,Rohit;Baron,Nofar;Gilad,Ayelet;Wagner,Gerhard;Léger-Abraham,Mélissa;Shapira,Michal
通讯作者：
Shapira,Michal