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 翻译控制是细胞中的主要基因表达调控机制之一 失调导致许多人类疾病。核糖体通常被视为本构分子 但是，发生蛋白质合成的机器，但是最近挑战了这种观点 核糖体专业化和开放全新研究领域的假设。这个项目将调查 使用利什曼原虫作为模型生物的原生动物中核糖体专业化的基本概念。在 与其他真核生物形成鲜明对比的是，包括利什曼原虫在内的锥虫是单细胞生物及其 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 出色的模型生物体研究核糖体专业在mRNA翻译调节中的作用。 包括温度，pH，营养条件在内的环境在基因表达调节中起着重要作用， 但是，鲜为人知的是，分子参与者参与了翻译调节 环境压力和主机的变化。众所周知，在热量期间翻译被全局压抑 震惊，但是，一些mRNA逃脱了翻译抑制及其翻译的增强。翻译 编码参与压力反应的蛋白质的mRNA对于Leishmania的应对能力非常重要 随着压力，其分化和生存，尚不清楚热诱导的mRNA如何逃脱 热应力期间的全球翻译压抑。该项目基于核糖体的假设 组合物在热应力期间发生实质性变化，以促进子集的有效翻译 编码参与压力反应的蛋白质的mRNA。拟议的研究将在 几个不同的级别：（目标1）它将在积极翻译的mRNA的全基因组量表子集上识别 在热应激期间； （AIM 2）它将揭示高热期间利什曼原虫的核糖体组成的变化 压力;最后，（目标3）它将使用CRISPR/CAS9敲除屏幕检查 热诱导的转录本的选择性翻译及其在利什曼尼亚生命周期中所起的作用。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/调节剂在原生动物的压力期间有效地翻译并确定其作用 在利什曼尼亚分化中。