BYPASS MECHANISMS IN EUKARYOTIC REPLICATION

真核复制中的旁路机制

基本信息

  • 批准号:
    10249266
  • 负责人:
  • 金额:
    $ 24.9万
  • 依托单位:
  • 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:
    2018-09-01 至 2023-08-31
  • 项目状态:
    已结题

项目摘要

PROJECT SUMMARY/ABSTRACT Chromosomes are copied by a complex holoenzyme called the replisome. Obstacles are routinely negotiated by the replisome with auxiliary mechanisms that ensure genomic integrity, aberrance of which can lead to chromosome instability and a broad range of diseases including cancer. The candidate’s long term goal is to understand the molecular basis for genetic and epigenetic fidelity, with the potential to improve the treatment and/or prevention of disease. In this proposal, the candidate will use a fully functional replisome reconstituted from over 30 pure polypeptides to study how replisomes bypass obstacles that regularly occur in the genome while enforcing genetic and epigenetic integrity across generations. In the first specific aim the candidate’s current work on the molecular mechanisms of lesion bypass by the replisome will be elaborated, with a focus on how checkpoint kinases Mec1 and Rad53 and the Mrc1/Tof1/Csm3 (MTC) complex modify the activity of the replisome while regulating lesion bypass. Interactions between several key replisome components and the MTC complex will be probed by microscale thermophoresis (MST) and cross-linking mass spectrometry (XL- MS). In addition to biochemical experiments, single-molecule approaches will be used to probe the mechanism of replisome regulation by the MTC complex, resolving replisome components with fluorescence during active replication on DNA. Single-molecule FRET experiments will be used in the independent phase to probe how MTC affects the structural dynamics of the replisome. In the second aim, the candidate will investigate nucleosome bypass by the replisome, focusing on the post-replication fate of histones during the mentored phase. Using histones enriched for fluorescence, replication-coupled histone deposition will be tracked by a first-of-its-kind attempt at spatially resolving leading vs. lagging strand products with a combination of optical trapping, fluorescence, and flow. Along with bead-based biochemical experiments, the results will help differentiate between models of epigenetic inheritance. In the independent phase, interactions between the replisome and FACT, a histone chaperone, will be determined with MST and XL-MS. The molecular mechanisms of various chaperones will be probed using single-molecule FRET experiments monitoring the spatiokinetics of chromatin remodeling in real time, also determining the role of histone modifications in remodeling. The mentored phase of the project will be conducted in the laboratories of Dr. Michael O’Donnell (mentor) and Dr. Shixin Liu (co-mentor) at Rockefeller University, a world-class research environment. The success of the candidate’s proposed research depends critically on using advanced integrative single-molecule techniques, as well as XL-MS. Thus, the candidate seeks intensive training with manipulation and detection of individual molecules in the co-mentor’s state-of-the-art facilities, in addition to XL-MS with Drs. Brian Chait and Yi Shi (collaborators). The candidate also has also planned activities to improve mentoring, lab management, scientific communication, and professional skills, enabling a successful transition to an independent career.
项目总结/摘要 染色体是由一种叫做复制体的复杂全酶复制的。障碍是例行谈判 通过复制体与确保基因组完整性的辅助机制,其变异可导致 染色体不稳定性和包括癌症在内的多种疾病。候选人的长期目标是 了解遗传和表观遗传保真度的分子基础,有可能改善治疗 和/或预防疾病。在这个提议中,候选人将使用一个功能齐全的复制体重组 从30多个纯多肽中研究复制体如何绕过基因组中经常发生的障碍 同时加强遗传和表观遗传的完整性。在第一个具体目标中, 目前的工作,病变旁路的分子机制的复制体将详细阐述,重点是 关于检查点激酶Mec 1和Rad 53以及Mrc 1/Tof 1/Csm 3(MTC)复合物如何改变 复制体,同时调节病变旁路。几个关键的复制体组分与 MTC复合物将通过微尺度热泳(MST)和交联质谱(XL-100)探测。 MS)。除了生化实验外,单分子方法将用于探索机制 MTC复合物对复制体的调节,在活性期间用荧光解析复制体组分, DNA复制单分子FRET实验将用于独立阶段,以探索如何 MTC影响复制体的结构动力学。在第二个目标中,候选人将调查 核小体旁路的复制体,集中在复制后的命运组蛋白在指导 相位使用荧光富集的组蛋白,复制偶联的组蛋白沉积将通过一种新的方法来跟踪。 第一次尝试在空间上解决领先与落后的链产品与光学组合 捕获荧光和流动沿着以珠子为基础的生化实验, 区分不同的表观遗传模型。在独立阶段, 将用MST和XL-MS测定复制体和FACT(一种组蛋白伴侣)的分子量。 各种分子伴侣的机制将使用单分子FRET实验监测 在真实的时间内染色质重塑的空间分布,也决定了组蛋白修饰在 重塑该项目的指导阶段将在Michael奥唐纳博士的实验室进行 他是洛克菲勒大学(Rockefeller University)的导师,刘世新博士(Shixin Liu)是他的共同导师,洛克菲勒大学拥有世界一流的研究环境。的 候选人提出的研究的成功关键取决于使用先进的整合单分子 技术,以及XL-MS。因此,候选人寻求与操纵和检测的密集培训, 在共同导师的国家的最先进的设施,除了XL-MS与布赖恩Chait博士和 李(合作者)。候选人还计划了一些活动,以改善指导,实验室管理, 科学沟通和专业技能,使成功过渡到一个独立的职业生涯。

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 }}

Grant Schauer其他文献

Grant Schauer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('Grant Schauer', 18)}}的其他基金

Bypass Mechanisms in Eukaryotic Replication
真核复制中的旁路机制
  • 批准号:
    10500889
  • 财政年份:
    2022
  • 资助金额:
    $ 24.9万
  • 项目类别:
Bypass Mechanisms in Eukaryotic Replication
真核复制中的旁路机制
  • 批准号:
    10672310
  • 财政年份:
    2022
  • 资助金额:
    $ 24.9万
  • 项目类别:
Bypass Mechanisms in Eukaryotic Replication
真核复制中的旁路机制
  • 批准号:
    10798784
  • 财政年份:
    2022
  • 资助金额:
    $ 24.9万
  • 项目类别:
BYPASS MECHANISMS IN EUKARYOTIC REPLICATION
真核复制中的旁路机制
  • 批准号:
    10017302
  • 财政年份:
    2018
  • 资助金额:
    $ 24.9万
  • 项目类别:

相似海外基金

Rational design of rapidly translatable, highly antigenic and novel recombinant immunogens to address deficiencies of current snakebite treatments
合理设计可快速翻译、高抗原性和新型重组免疫原,以解决当前蛇咬伤治疗的缺陷
  • 批准号:
    MR/S03398X/2
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Fellowship
Re-thinking drug nanocrystals as highly loaded vectors to address key unmet therapeutic challenges
重新思考药物纳米晶体作为高负载载体以解决关键的未满足的治疗挑战
  • 批准号:
    EP/Y001486/1
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Research Grant
CAREER: FEAST (Food Ecosystems And circularity for Sustainable Transformation) framework to address Hidden Hunger
职业:FEAST(食品生态系统和可持续转型循环)框架解决隐性饥饿
  • 批准号:
    2338423
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Continuing Grant
Metrology to address ion suppression in multimodal mass spectrometry imaging with application in oncology
计量学解决多模态质谱成像中的离子抑制问题及其在肿瘤学中的应用
  • 批准号:
    MR/X03657X/1
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Fellowship
CRII: SHF: A Novel Address Translation Architecture for Virtualized Clouds
CRII:SHF:一种用于虚拟化云的新型地址转换架构
  • 批准号:
    2348066
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Standard Grant
BIORETS: Convergence Research Experiences for Teachers in Synthetic and Systems Biology to Address Challenges in Food, Health, Energy, and Environment
BIORETS:合成和系统生物学教师的融合研究经验,以应对食品、健康、能源和环境方面的挑战
  • 批准号:
    2341402
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Standard Grant
The Abundance Project: Enhancing Cultural & Green Inclusion in Social Prescribing in Southwest London to Address Ethnic Inequalities in Mental Health
丰富项目:增强文化
  • 批准号:
    AH/Z505481/1
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Research Grant
ERAMET - Ecosystem for rapid adoption of modelling and simulation METhods to address regulatory needs in the development of orphan and paediatric medicines
ERAMET - 快速采用建模和模拟方法的生态系统,以满足孤儿药和儿科药物开发中的监管需求
  • 批准号:
    10107647
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    EU-Funded
Ecosystem for rapid adoption of modelling and simulation METhods to address regulatory needs in the development of orphan and paediatric medicines
快速采用建模和模拟方法的生态系统,以满足孤儿药和儿科药物开发中的监管需求
  • 批准号:
    10106221
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    EU-Funded
Recite: Building Research by Communities to Address Inequities through Expression
背诵:社区开展研究,通过表达解决不平等问题
  • 批准号:
    AH/Z505341/1
  • 财政年份:
    2024
  • 资助金额:
    $ 24.9万
  • 项目类别:
    Research Grant
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了