The Mechanism of Elimination of the Mitochondrial DNA Replisome

线粒体DNA复制体的消除机制

基本信息

项目摘要

Abstract Defects in the human mitochondrial DNA (mtDNA) replication process result in the accumulation of mutations that give rise to a broad spectrum of degenerative disorders involving multiple organs and systems of the human body. In this project, we propose to investigate a putative mechanism that prevents the formation of large-scale deletions in mtDNA, which are the most common (de novo) defects of the mitochondrial genome. The mechanism of deletion formation is unknown, but studies reported to date indicate that they originate from stalling of the mitochondrial DNA replication machinery (the replisome), which leads to the breaking of DNA strands. Notably, prominent mtDNA replication stalling sites correspond with binding sites of the major mitochondrial protease, Lon, which suggests that it might be involved in the elimination of halted replisomes. In addition, the components of human mitochondrial Hsp70/40 chaperone system have been found to co-precipitate with the catalytic subunit of the mitochondrial replicative polymerase, which suggests that these may be involved in the assembly or disassembly of mitochondrial replisomes. In fact, in model organisms, Hsp70/40 systems have been found to assist Lon protease by unfolding and delivering protein substrates. On the other hand, Hsp70/40 systems have also been found to cooperate with another protease, ClpXP, which deficiency results in mitochondrial genome destabilization. This proposal aims to evaluate the hypothesis that stalled mtDNA replisomes are eliminated by two alternative mechanisms that engage either Lon or ClpXP protease. In addition, the Hsp70/40 chaperone system may serve to disassemble the replisome and deliver its components to the client protease. Confirmation and characterization of a direct relationship between the capacity of a cell to remove defective mitochondrial replisomes and the accumulation of damage in the mitochondrial genome, would bring a novel and exciting perspective on the development of mtDNA deletions-linked human disorders, potentially identifying targets for prospective therapeutic strategies. To this end, we will apply a comprehensive approach combining the cutting- edge technique of biolayer interferometry for the analysis of molecular affinities and kinetic parameters, a methodical biochemical analysis that entails specialized enzymatic assays, and testing the putative correlation between cellular levels of Lon, Hsp70/40 and ClpX and the development of mtDNA deletions in vivo, using Saccharomyces cerevisiae as the model organism. To date, we have demonstrated that Lon protease hydrolases the catalytic subunit of the mitochondrial replicative polymerase, but only in the absence of the remaining polymerase subunits. This finding confirms our initial hypothesis and warrants the investigation of the role of Hsp70/40 and/or ClpX in the disassembly of the replisome complex to enable degradation of the catalytic core by Lon protease.
摘要 人类线粒体DNA(mtDNA)复制过程中的缺陷导致 突变引起广泛的退行性疾病,涉及多个器官和系统, 人体的在这个项目中,我们建议调查一个假定的机制,防止形成 线粒体DNA中的大规模缺失,这是线粒体基因组中最常见的(从头)缺陷。 缺失形成的机制尚不清楚,但迄今报道的研究表明,它们起源于 线粒体DNA复制机制(复制体)停滞,导致DNA断裂 股。值得注意的是,突出的mtDNA复制停滞位点对应于主要的线粒体DNA的结合位点。 线粒体蛋白酶,Lon,这表明它可能参与停止复制体的消除。在 此外,已发现人线粒体Hsp 70/40分子伴侣系统的组分共沉淀 与线粒体复制聚合酶的催化亚基,这表明这些可能涉及 在线粒体复制体的组装或拆卸中。事实上,在模式生物中,Hsp 70/40系统具有 已发现通过解折叠和递送蛋白质底物来辅助Lon蛋白酶。Hsp70/40 还发现系统与另一种蛋白酶ClpXP协作,其缺陷导致 线粒体基因组不稳定。 该提议旨在评估停滞的mtDNA复制体被两个 参与Lon或ClpXP蛋白酶的替代机制。此外,Hsp 70/40分子伴侣系统 可用于分解复制体并将其组分递送至客户蛋白酶。确认和 表征细胞去除缺陷线粒体的能力之间的直接关系 复制体和线粒体基因组中损伤的积累,将带来一个新的和令人兴奋的 mtDNA缺失相关人类疾病的发展前景,可能确定目标, 前瞻性治疗策略。为此,我们将采取综合办法, 用于分子亲和力和动力学参数分析的生物膜干涉边缘技术 系统的生化分析,需要专门的酶测定,并测试推定的相关性 在细胞水平的Lon,Hsp 70/40和ClpX与体内mtDNA缺失的发展之间,使用 酿酒酵母作为模式生物。到目前为止,我们已经证明了Lon蛋白酶水解酶 线粒体复制聚合酶的催化亚基,但仅在缺乏其余亚基的情况下 聚合酶亚单位。这一发现证实了我们最初的假设,并保证调查的作用 Hsp 70/40和/或ClpX在复制体复合物的分解中的作用,以使催化核心能够降解 Lon蛋白酶。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Grzegorz Leszek Ciesielski其他文献

Grzegorz Leszek Ciesielski的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Grzegorz Leszek Ciesielski', 18)}}的其他基金

The mechanism of elimination of the mitochondrial DNA replisome
线粒体DNA复制体的消除机制
  • 批准号:
    10880042
  • 财政年份:
    2021
  • 资助金额:
    $ 20.99万
  • 项目类别:
The Mechanism of Elimination of the Mitochondrial DNA Replisome
线粒体DNA复制体的消除机制
  • 批准号:
    10582403
  • 财政年份:
    2021
  • 资助金额:
    $ 20.99万
  • 项目类别:

相似海外基金

Adenosine triphosphate as a master variable for biomass in the oceanographic context
三磷酸腺苷作为海洋学背景下生物量的主变量
  • 批准号:
    2319114
  • 财政年份:
    2023
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Standard Grant
Characterizing the Interaction Between Adenosine Triphosphate and Pathological Alpha-synuclein Structures in Parkinson's Disease
表征帕金森病中三磷酸腺苷与病理性 α-突触核蛋白结构之间的相互作用
  • 批准号:
    565727-2021
  • 财政年份:
    2021
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Alexander Graham Bell Canada Graduate Scholarships - Master's
Investigation of the development of pain during orthodontic tooth movement with adenosine triphosphate
三磷酸腺苷正畸牙齿移动过程中疼痛发生的研究
  • 批准号:
    20K18789
  • 财政年份:
    2020
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Neural Regulation of Adenosine Triphosphate (ATP) in the Nasal Mucosa
鼻粘膜三磷酸腺苷 (ATP) 的神经调节
  • 批准号:
    19K18793
  • 财政年份:
    2019
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Dynamics of the oxygen-dependent release of adenosine triphosphate from erythrocytes
红细胞氧依赖性三磷酸腺苷释放的动力学
  • 批准号:
    460605-2014
  • 财政年份:
    2016
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Alexander Graham Bell Canada Graduate Scholarships - Doctoral
Development of an Analytical Tool Utilizing Electrochemical Detection Methods For the Measuring of Protein Kinase Activity on a Protein Substrate Using Ferrocene-Adenosine Triphosphate (Fc-ATP) as a C
利用电化学检测方法开发分析工具,以二茂铁-三磷酸腺苷 (Fc-ATP) 作为 C,测量蛋白质底物上的蛋白激酶活性
  • 批准号:
    469948-2014
  • 财政年份:
    2016
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Vanier Canada Graduate Scholarship Tri-Council - Doctoral 3 years
Adenosine Triphosphate as a Signal for Evaluating Microbial Risk from Groundwater Supplies
三磷酸腺苷作为评估地下水供应微生物风险的信号
  • 批准号:
    507411-2016
  • 财政年份:
    2016
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Engage Grants Program
Development of an Analytical Tool Utilizing Electrochemical Detection Methods For the Measuring of Protein Kinase Activity on a Protein Substrate Using Ferrocene-Adenosine Triphosphate (Fc-ATP) as a C
利用电化学检测方法开发分析工具,以二茂铁-三磷酸腺苷 (Fc-ATP) 作为 C,测量蛋白质底物上的蛋白激酶活性
  • 批准号:
    469948-2014
  • 财政年份:
    2015
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Vanier Canada Graduate Scholarship Tri-Council - Doctoral 3 years
Dynamics of the oxygen-dependent release of adenosine triphosphate from erythrocytes
红细胞氧依赖性三磷酸腺苷释放的动力学
  • 批准号:
    460605-2014
  • 财政年份:
    2015
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Alexander Graham Bell Canada Graduate Scholarships - Doctoral
Dynamics of the oxygen-dependent release of adenosine triphosphate from erythrocytes
红细胞氧依赖性三磷酸腺苷释放的动力学
  • 批准号:
    460605-2014
  • 财政年份:
    2014
  • 资助金额:
    $ 20.99万
  • 项目类别:
    Alexander Graham Bell Canada Graduate Scholarships - Doctoral
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了