Biogenesis of macromolecular machines for post-transcriptional regulation of translation

用于翻译转录后调控的大分子机器的生物发生

基本信息

批准号：
10798922
负责人：
Homa Ghalei
金额：
$ 12.1万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-17 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10798922
关键词：
ATP phosphohydrolase Address Affect Autoimmune Diseases Award Biochemical Biochemistry Biogenesis Biological Assay Biology Cell physiology Cells Cellular Stress Colon Carcinoma Colorectal Cancer Complex Coupled Cues Data Defect Deposition Development Disease Down-Regulation Drug resistance Edema Elements Encephalopathies Ensure Eukaryota Event Foundations Funding Future Gene Expression Genetic Genetic Transcription Genome Goals Guide RNA Human Hypsarrhythmia Individual Isomerism Knowledge Laboratories Lead Link Malignant Neoplasms Malignant neoplasm of brain Malignant neoplasm of lung Malignant neoplasm of prostate Maps Mediating Methylation Modification Molecular Morphology Mutation National Institute of General Medical Sciences Nerve Degeneration Nucleotides Optic Atrophy Output Pathologic Pathway interactions Pattern Perception Play Post-Transcriptional Regulation Prader-Willi Syndrome Process Production Protein Biosynthesis Proteins Proteomics Pseudouridine Public Health RNA RNA Splicing Reagent Regulation Regulatory Pathway Research Research Personnel Ribonucleases Ribonucleoproteins Ribosomal RNA Ribosomes Role Site Small Nucleolar RNA Spliceosomes Stress Syndrome Textbooks Therapeutic Intervention Transcription Alteration Translational Regulation Translations Untranslated RNA Uridine Yeasts Zebrafish cancer type career cell growth falls human disease insight malignant breast neoplasm nervous system disorder new therapeutic target next generation sequencing novel posttranscriptional programs protein complex structural biology tool translational impact tumorigenesis yeast genetics

项目摘要

Summary: Ribosomes are highly conserved RNA-protein complexes that direct protein synthesis in all cells. Dysregulation of ribosome production or function is detrimental to gene expression and underlies several disease states. Over 2% of ribosomal RNA (rRNA) nucleotides are modified. These modifications play a critical role in the proper production of ribosomes that can accurately perform protein synthesis. The two major rRNA modifications are 2’-O-methylation and pseudouridylation that are directed by highly conserved non-coding RNAs called small nucleolar RNAs (snoRNAs). Altered levels of snoRNAs are associated with human diseases from neurodegeneration to multiple types of cancer, underscoring their importance for proper cell growth. Therefore, a key question is how levels of snoRNAs are regulated and how does their dysregulation lead to translation defects in disease? Despite the textbook perception that rRNA modifications are equally deposited in all ribosomes, recent advances in mapping modifications have revealed substoichiometric rRNA modification sites, strongly suggesting that ribosome assembly and function may be regulated by the modification status of rRNA. A long-term goal of my laboratory is to identify the post-transcriptional mechanisms that regulate the abundance A prominent rRNA modification in eukaryotes is 2’-O-methylation, the incorporation of which is guided by snoRNAs of the box C/D class. These snoRNAs interact with a set of evolutionarily conserved proteins to form ribonucleoprotein complexes (snoRNPs). The assembly of snoRNPs is highly regulated which, in turn, is important to maintain levels of snoRNAs and to coordinate this process with other cellular events. However, despite their fundamental importance, much of these regulatory events remains a black box. We have performed targeted yeast mutational and suppressor screens of snoRNP assembly factors to determine their essential contributions and identify genetic pathways that mediate snoRNP biogenesis. Our data indicate that regulation of box C/D snoRNP production by assembly factors is critically important for control of the modification pattern of rRNAs and dysregulation of this process alters the biogenesis pathway and the fidelity of ribosomes. Our goal is to combine the novel genetic tools and reagents that we have recently developed with to answer two key questions: 1) regulatory factors control the steady-state levels of snoRNAs required for accurate modification of rRNA?; and 2) How do changes in snoRNA levels alter and tune protein synthesis? These studies will provide significant insights into the control of gene expression by snoRNAs at the translation level, and may inform our view of how snoRNA dysregulation underlies human disease. of snoRNAs and understand their contribution to cellular translational control. biochemical assays, structural biology, proteomics, and next-generation sequencing How do

概括：核糖体是高度保守的RNA-蛋白质复合物，可导致所有细胞中的蛋白质合成。失调核糖体的产生或功能对基因表达有害，并且是几种疾病状态的基础。超过 2％的核糖体RNA（RRNA）核动肽被修饰。这些修改在适当的可以准确执行蛋白质合成的核糖体的产生。两个主要的rRNA修改是由高度组成的非编码RNA指导的2'-O-甲基化和假硫岛化称为小核仁RNA（snornas）。 snornas水平的改变与人类疾病有关神经变性到多种类型的癌症，强调了它们对适当细胞生长的重要性。所以，一个关键的问题是如何调节snornas的水平，以及它们的失调如何导致翻译疾病缺陷？尽管教科书认为rRNA修改同样沉积在所有人中核糖体，映射修饰的最新进展揭示了化学计量rRNA修饰位点，强烈表明核糖体组装和功能可以通过rRNA的修饰状态来调节。我实验室的长期目标是确定调节抽象的转录后机制突出的rRNA修改真核生物是2'-O-甲基化，其掺入的融合是由盒子C/D类的snornas指导的。这些 SNORNAS与一组进化的蛋白质相互作用，形成核糖核蛋白复合物（snornps）。 snornps的组装受到高度调节，而这对于维持水平很重要 snornas并与其他细胞事件进行协调。但是，使他们的基本任务重要的是，这些监管事件中的大部分仍然是黑匣子。我们已经进行了靶向酵母突变和snornp装配因素的抑制屏幕以确定其基本贡献并确定介导SNORNP生物发生的遗传途径。我们的数据表明Box C/D SNORNP的调节通过组装因素生产对于控制RRNA的修饰模式至关重要该过程的失调改变了核糖体的生物发生途径和忠诚度。我们的目标是结合我们最近开发的新型遗传工具和试剂回答两个关键问题：1）监管因素控制精确修饰rRNA所需的snornas的稳态水平？ 2）如何改变 SNORNA水平改变并调整蛋白质合成？这些研究将为控制 snornas在翻译层面的基因表达，并可能告知我们对snorna失调的看法是人类疾病的基础。 snornas，并了解它们对细胞翻译控制的贡献。生化测定，结构生物学，蛋白质组学和下一代测序如何

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RNA structural probing of guanine and uracil nucleotides in yeast.

DOI：
10.1371/journal.pone.0288070
发表时间：
2023
期刊：
PloS one
影响因子：
3.7
作者：
通讯作者：

A Biallelic Variant of the RNA Exosome Gene EXOSC4 Causes Translational Defects Associated with a Neurodevelopmental Disorder.

RNA 外泌体基因 EXOSC4 的双等位变体会导致与神经发育障碍相关的翻译缺陷。

DOI：
10.1101/2023.10.24.23297197
发表时间：
2023
期刊：
medRxiv : the preprint server for health sciences
影响因子：
0
作者：
Fasken,MiloB;Leung,SaraW;Cureton,LaurynA;Al-Awadi,Maha;Al-Kindy,Adila;Khoshnevis,Sohail;Ghalei,Homa;Al-Maawali,Almundher;Corbett,AnitaH
通讯作者：
Corbett,AnitaH

Practical advice for mentoring and supporting faculty colleagues in STEM fields: Views from mentor and mentee perspectives.

DOI：
10.1016/j.jbc.2021.101062
发表时间：
2021-09
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Spangle JM;Ghalei H;Corbett AH
通讯作者：
Corbett AH

Ribosomal RNA 2'-O-methylations regulate translation by impacting ribosome dynamics.

DOI：
10.1073/pnas.2117334119
发表时间：
2022-03-22
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Khoshnevis S;Dreggors-Walker RE;Marchand V;Motorin Y;Ghalei H
通讯作者：
Ghalei H

Studies of mutations of assembly factor Hit1 in budding yeast suggest translation defects as the molecular basis for PEHO syndrome.

DOI：
10.1016/j.jbc.2022.102261
发表时间：
2022-09
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Dreggors-Walker, R. Elizabeth;Cohen, Lauren N.;Khoshnevis, Sohail;Marchand, Virginie;Motorin, Yuri;Ghalei, Homa
通讯作者：
Ghalei, Homa