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

• 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
• 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

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.

项目摘要 翻译调控是细胞中主要的基因表达调控机制之一， 失调导致许多人类疾病。核糖体一般被认为是组成性分子 然而，这种观点最近受到了挑战，支持了 核糖体特化假说，开辟了一个全新的研究领域。本项目将调查 以利什曼原虫为模式生物，阐述了原生动物核糖体特化的基本概念。在 与其他真核生物不同，包括利什曼原虫属物种在内的锥虫是单细胞生物， 基因表达的控制主要在mRNA翻译期间实现。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极好模式生物。 环境包括温度、pH、营养条件对基因表达调控起着重要作用， 然而，人们对哪些分子参与了翻译过程中的调控知之甚少。 环境压力和宿主的变化。众所周知，在高温下， 然而，在休克时，一些mRNA逃脱了翻译抑制，它们的翻译被增强。翻译 编码参与应激反应的蛋白质的mRNA对利什曼原虫科普能力非常重要 然而，对于热诱导的mRNA是如何逃逸的， 热应激过程中的整体翻译抑制。这个项目是基于核糖体 组合物在热应力期间经历实质性变化，以促进亚组的有效翻译。 编码参与应激反应的蛋白质的mRNA。拟议的研究将提供新的资料， 几个不同的水平：（目标1）它将在全基因组范围内识别积极翻译的mRNA子集 目的2：揭示热应激过程中利什曼原虫核糖体组成的变化 最后，（目标3）它将使用CRISPR/Cas9敲除筛选来检查哪些蛋白质确实促进 热诱导转录的选择性翻译以及它们在利什曼原虫的生命周期中扮演的角色。这 综合多学科的方法将首次揭示成绩单如何选择性地依赖于特定的 核糖体组分/调节剂在原生动物应激期间的有效翻译，并确定其作用 利什曼原虫的分化