Ribosome remodeling as a mechanism of translational control during stress

核糖体重塑作为应激期间翻译控制的机制

基本信息

批准号：
10435068
负责人：
Jorge Salazar-Bravo
金额：
$ 45.9万
依托单位：
TEXAS TECH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10435068
关键词：
Affect Animal Model Bioinformatics CRISPR/Cas technology Cells Complement Data Disease Down-Regulation Ensure Environment Eukaryota Evaluation Exposure to Gene Expression Gene Expression Regulation Genes Genetic Translation Heat Stress Disorders Heat shock proteins Heat-Shock Response Human Infection Insecta Knock-out Leishmania Leishmania major Leishmaniasis Life Cycle Stages Light Messenger RNA Modeling Molecular Molecular Machines Organism Parasites Persons Play Polyribosomes Protein Analysis Protein Biosynthesis Proteins Proteomics Protozoa RNA RNA Interference RNA-Binding Proteins Regulation Research Ribosomal Proteins Ribosomes Role Stress Techniques Temperature Testing Time Transcript Transcriptional Regulation Translating Translational Repression Translations Up-Regulation Western Blotting base biological adaptation to stress comparative environmental change environmental stressor experimental study fighting genome-wide human disease interdisciplinary approach mRNA Stability next generation sequencing novel nutrition polysome profiling protein expression response ribosome profiling stoichiometry

项目摘要

PROJECT SUMMARY Translational control is one of the major gene expression regulation mechanisms in the cell and its dysregulation leads to many human diseases. Ribosomes in general are viewed as constitutive molecular machines where protein synthesis takes place, however, this view has been recently challenged supporting the hypothesis of ribosome specialization and opening completely new field of research. This project will investigate the fundamental concept of ribosome specialization in protozoa using Leishmania as a model organism. In contrast to other eukaryotes, trypanosomatids including Leishmania species are unicellular organisms and their control of gene expression is mostly achieved during mRNA translation. Therefore, this protozoan represents an excellent model organism to study the role of ribosome specialization in mRNA translation regulation. Environment including temperature, pH, nutrition conditions plays a big role in gene expression regulation, however, it is poorly understood what molecular players are involved in the regulation of translation during environmental stresses and change of host. It is known that translation is globally repressed during the heat shock, however, some mRNAs escape translational repression and their translation is enhanced. Translation of mRNAs encoding for proteins involved in stress response is very important for the Leishmania ability to cope with stress, its differentiation and survival, however, it is not well understood how heat-induced mRNAs escape the global translational repression during the heat stress. This project is based on the hypothesis that ribosome composition undergoes a substantial change during heat stress to promote efficient translation of subset of mRNAs encoding for proteins involved in stress response. The proposed study will provide new information at several different levels: (Aim 1) it will identify on genome-wide scale subset of mRNAs that are actively translated during the heat stress; (Aim 2) it will reveal changes in ribosome composition of Leishmania during the heat stress; and finally, (Aim 3) it will examine using CRISPR/Cas9 knock-out screen what proteins indeed promote selective translation of heat-induced transcripts and what role they play in the life cycle of Leishmania. This comprehensive multidisciplinary approach will reveal for the first time how transcripts selectively rely on specific ribosome components/regulators for their efficient translation during stress in protozoa and establish their role in Leishmania differentiation.

项目概要翻译控制是细胞及其基因表达调控的主要机制之一失调会导致许多人类疾病。核糖体通常被视为组成分子然而，这种观点最近受到了挑战，支持核糖体特化假说并开辟了全新的研究领域。本项目将调查使用利什曼原虫作为模式生物的原生动物核糖体特化的基本概念。在与其他真核生物相比，包括利什曼原虫在内的锥虫是单细胞生物，其基因表达的控制主要是在 mRNA 翻译过程中实现的。因此，这种原生动物代表了研究核糖体特化在 mRNA 翻译调控中的作用的优秀模型生物。环境包括温度、pH、营养条件对基因表达调控起着很大的作用，然而，人们对哪些分子参与者参与翻译调控知之甚少。环境压力和宿主的变化。众所周知，在炎热的天气里，翻译会受到全局抑制。然而，令人震惊的是，一些 mRNA 逃脱了翻译抑制，它们的翻译得到了增强。请先登录再添加翻译编码参与应激反应的蛋白质的 mRNA 对于利什曼原虫的应对能力非常重要然而，热诱导的 mRNA 如何逃逸尚不清楚热应激期间的整体平移抑制。该项目基于核糖体的假设在热应激期间，其组成发生了实质性变化，以促进子集的有效翻译编码参与应激反应的蛋白质的 mRNA。拟议的研究将提供新信息几个不同的级别：（目标 1）它将在全基因组规模上识别正在积极翻译的 mRNA 子集热应激期间；（目标 2）它将揭示利什曼原虫在高温期间核糖体组成的变化压力;最后，（目标 3）它将使用 CRISPR/Cas9 敲除筛选检查蛋白质确实促进什么热诱导转录物的选择性翻译及其在利什曼原虫生命周期中发挥的作用。这全面的多学科方法将首次揭示成绩单如何选择性地依赖特定的核糖体成分/调节因子在原生动物应激期间有效翻译并确定其作用利什曼原虫分化。