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Molecular mechanisms of mtDNA inheritance

线粒体DNA遗传的分子机制

基本信息

批准号：
10371889
负责人：
Samantha C Lewis
金额：
$ 24.9万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-07 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10371889
关键词：
ATP Synthesis Pathway Acute Address Alleles Attenuated Bar Codes Biology Buffers CRISPR interference Cell physiology Cells Cellular biology Circular DNA Critical Pathways DNA DNA Maintenance DNA biosynthesis DNA copy number Data Disease Endoplasmic Reticulum Escherichia coli Etiology Eukaryotic Cell Event Fostering Gene Cluster Genes Genetic Genetic Transcription Genome Goals Growth Haplotypes Heritability High-Throughput Nucleotide Sequencing Human Individual Laboratories Lead Libraries Link Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Mediating Metabolic Diseases Microscopy Microtubules Mission Mitochondria Mitochondrial DNA Mitochondrial Diseases Mitochondrial Inheritance Mitochondrial Respiratory Chain Deficiencies Molecular Mutation Nerve Degeneration Organelles Oxidative Phosphorylation Pathway interactions Postdoctoral Fellow Process Proteins Proteomics RNA Recovery Regulation Research Research Personnel Residual state Role Science Severity of illness Site Structure System Technology Testing Tissues Topoisomerase Topoisomerase III United States National Institutes of Health Viral Work Zalcitabine antiviral nucleoside analog base cell growth cell motility cell type cohort comparative constriction disease phenotype experimental study genome wide screen genome-wide human disease insight knock-down live cell microscopy mitochondrial DNA mutation mitochondrial genome mutant new therapeutic target novel nuclease professor respiratory response segregation single molecule real time sequencing single-cell RNA sequencing targeted treatment transcriptome sequencing

项目摘要

Project Summary Mitochondrial DNA (mtDNA) encodes RNAs and proteins critical for cell function. However, pathways that control mtDNA segregation and copynumber in mammalian cells are not well understood. The goals of this work are to identify and characterize the unknown protein machinery that segregates replicating mtDNAs into discrete nucleoid structures at ER-mitochondria contact sites and to describe the mechanisms and pathways that regulate mtDNA copy number. The proposed experiments will provide fundamental insight into the mechanism of mtDNA segregation and copynumber control, and how those processes are regulated, potentially leading to the discovery and characterization of novel pathways that regulate the inheritance of mtDNA disease haplotypes. In Aim 1, cutting-edge live-cell microscopy, proteomics, and high-throughput sequencing technologies will be employed to dissect the molecular functions of mtDNA segrosome candidate proteins identified in preliminary experiments. Aim 2 will address which cellular pathways are critical to regulation of mtDNA copy number in human cells. This will be accomplished by systematic genome-wide screens for modulators of mtDNA depletion recovery, single-cell RNA sequencing to cluster genes by their transcriptional responses to mtDNA loss, and the characterization of candidate mtDNA copynumber effector proteins in human cells depleted of mtDNA. These experiments will provide fundamental insight into the mechanism of mtDNA segregation and copynumber control, and how those processes are regulated, potentially leading to the discovery and characterization of novel pathways that regulate the inheritance of mtDNA disease haplotypes.

项目摘要线粒体DNA(MtDNA)编码对细胞功能至关重要的RNA和蛋白质。然而，那些哺乳动物细胞中线粒体DNA的控制分离和拷贝数还不是很清楚。这样做的目的是工作是识别和表征未知的蛋白质机制，该机制将复制的mtDNA分离到内质网-线粒体接触部位的离散类核结构及其机制和途径来调节线粒体DNA拷贝数。拟议中的实验将提供对线粒体DNA分离和拷贝数控制的机制，以及这些过程是如何调控的，潜在地导致发现和表征调控遗传的新途径线粒体DNA病单倍型。在目标1中，尖端活细胞显微镜、蛋白质组学和高通量测序技术将被用来剖析mtdna segroome候选分子的功能。在初步实验中确定的蛋白质。目标2将解决哪些细胞通路对人类细胞线粒体DNA拷贝数的调控。这将通过系统化的全基因组完成筛选线粒体DNA耗竭恢复的调节子，单细胞RNA测序通过其基因簇线粒体DNA缺失的转录反应和候选线粒体DNA拷贝数效应因子的特征人类细胞中的蛋白质耗尽了线粒体DNA。这些实验将从根本上洞察线粒体DNA分离和拷贝数控制的机制，以及这些过程是如何调控的，潜在地导致发现和表征调控遗传的新途径线粒体DNA病单倍型。