Elucidating Molecular Mechanisms of Cancer Development by Investigating Key DNA Repair Pathways
通过研究关键 DNA 修复途径阐明癌症发展的分子机制
基本信息
- 批准号:10264134
- 负责人:
- 金额:$ 13.16万
- 依托单位:
- 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:2020-09-01 至 2023-08-31
- 项目状态:已结题
- 来源:
- 关键词:ATP phosphohydrolaseAmino AcidsAtomic Force MicroscopyBase PairingBindingBiochemicalBiochemistryBiological SciencesBiophysicsCancer EtiologyCell physiologyCellsChemistryCodeColorectal CancerComplementComplexConfocal MicroscopyCoupledDNADNA BindingDNA RepairDNA Repair GeneDNA Repair PathwayDNA biosynthesisDNA-Protein InteractionDataDaughterDetectionDevelopmentDiseaseDoctor of PhilosophyExonucleaseFailureFilmFluorescenceFluorescence Resonance Energy TransferFoundationsFutureGenerationsGenesGenomeGoalsHereditary Nonpolyposis Colorectal NeoplasmsHumanHuman GenomeHydrolysisIn VitroInheritedInstitutionKnowledgeLabelLeadLeftLinkMLH1 geneMalignant NeoplasmsMismatch RepairMismatch Repair DeficiencyMolecularMolecular ConformationMutationNobel PrizeNucleotidesOrganismParentsPathway interactionsPhysical ChemistryPhysicsPolymerasePositioning AttributePreparationProcessPropertyProteinsQuantum DotsResearchResearch PersonnelResearch TrainingScienceSignal TransductionSpectrum AnalysisStructureSystemTechniquesTestingTimeTissuesVisualizationWorkYeastsadvanced diseasebasecareerdesignelectric fieldexperienceexperimental studyin vivointerfacialmolecular modelingmutantnanoscalenanosciencenew therapeutic targetplasmonicsrepairedsingle moleculesingle-molecule FRETskillstenure tracktherapeutic targettumortumorigenesis
项目摘要
Project Summary
!
Dr. Sharonda LeBlanc earned her PhD in Nanoscale Science, investigating the effects of electric fields on
single quantum dot (SQD) fluorescence emission using confocal microscopy coupled with time-resolved
spectroscopy. She discovered interesting photophysical effects of moderate applied fields and plasmonic films
on SQDs. Dr. LeBlanc is currently transitioning from fundamental physical chemistry to biochemistry/biophysics
while still utilizing single molecule techniques. Currently, she is a postdoctoral researcher at UNC Chapel Hill
investigating molecular interactions of mismatch repair proteins. She works between two labs at UNC
(Chemistry) and NC State (Physics) to complete her research. Her career goal is to obtain a tenure-track
position at a research institution. She would like to combine her past research experience in nanoscience with
biochemistry/biophysics to ultimately investigate molecular mechanisms of cancer development in vivo. The
specific aims of this proposal and research training plan are designed to enhance Dr. LeBlanc's skills and
knowledge in the biological sciences, specifically DNA repair. Specific Aims 1 and 2 are as follows:
Specific Aim 1: Characterize the nucleotide-dependent dynamics of MutL conformations in the
absence of mismatch DNA in vitro using single molecule FRET.
Specific Aim 2: Investigate the dynamics of wild-type and mutant MutL conformations in the context of
mismatch repair initiation with nucleotides, MutS, and mismatch DNA in vitro with smFRET.
DNA mismatch repair (MMR) is a post-replicative system of proteins that corrects rare mistakes in the genome
of all organisms. In the human genome of 6 billion bases, there are ~ 600 errors per round of replication, per
cell. If left uncorrected, errors accumulate as permanent mutations in a genome, and can lead to a disease
state in the organism. MutS and MutL homologs are tasked with recognizing a mismatch in 107 correctly paired
bases, discriminating between parent and daughter strand, then initiating repair. Single amino acid mutations
in MutS and MutL proteins have been linked to hereditary and sporadic colorectal cancer, the third most
common cancer worldwide. Although these mutations, mostly associated with MutL, have been identified in
cancer cases, it is unclear how MMR deficiencies initiate and advance the disease. Failures in the mismatch
repair pathway likely initiate tumorigenesis, but we lack a fundamental understanding of the MMR process.
Single molecule fluorescence resonance energy transfer (smFRET) is uniquely capable of investigating the
molecular mechanism of MMR that involves multiple transient protein-protein and protein-DNA interactions.
These experiments may provide a basis for identifying therapeutic targets. The final aim of this proposal is
designed to facilitate Dr. LeBlanc's transition to an independent career, outlined in the Research Strategy:
Specific Aim 3: Develop ideas, design experiments, and test new hypotheses related to outstanding
questions in DNA repair in preparation for an independent research career.
项目摘要
!
博士Sharonda LeBlanc获得了纳米科学博士学位,研究电场对
单量子点(SQD)荧光发射使用共焦显微镜耦合时间分辨
谱她发现了适度应用场和等离子体薄膜的有趣物理效应
关于SQD LeBlanc博士目前正在从基础物理化学过渡到生物化学/生物物理学
同时仍然使用单分子技术。目前,她是美国查佩尔山的博士后研究员
研究错配修复蛋白的分子相互作用。她在两个实验室之间工作,
(化学)和NC状态(物理)完成她的研究。她的职业目标是获得终身教职
在一家研究机构工作。她想把她过去在纳米科学方面的研究经验与联合收割机结合起来,
生物化学/生物物理学,以最终研究体内癌症发展的分子机制。的
本提案和研究培训计划的具体目标旨在提高LeBlanc博士的技能,
生物科学知识,特别是DNA修复。具体目标1和2如下:
具体目标1:表征在细胞中MutL构象的核苷酸依赖性动力学
使用单分子FRET在体外不存在错配DNA。
具体目标2:研究野生型和突变型MutL构象的动力学,
用核苷酸、MutS和错配DNA在体外用smFRET启动错配修复。
DNA错配修复(MMR)是一种蛋白质复制后系统,可纠正基因组中的罕见错误
所有生物体的基因。在60亿个碱基的人类基因组中,每轮复制大约有600个错误,
cell.如果不加以纠正,错误就会积累成基因组中的永久性突变,并可能导致疾病
在有机体中。MutS和MutL同源物的任务是识别107个正确配对的错配
碱基,区分亲本和子链,然后启动修复。单个氨基酸突变
MutS和MutL蛋白与遗传性和散发性结直肠癌有关,这是第三大
全球常见癌症虽然这些突变,主要与MutL相关,已被确定在
在癌症病例中,目前还不清楚MMR缺陷如何启动和促进疾病。不匹配故障
修复途径可能启动肿瘤发生,但我们缺乏对MMR过程的基本了解。
单分子荧光共振能量转移(smFRET)是唯一能够研究
MMR的分子机制涉及多种瞬时蛋白质-蛋白质和蛋白质-DNA相互作用。
这些实验可能为识别治疗靶点提供基础。本提案的最终目的是
旨在促进LeBlanc博士向独立职业的过渡,如研究策略所述:
具体目标3:提出想法,设计实验,并测试与杰出的
DNA修复的问题,为独立的研究生涯做准备。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Sharonda LeBlanc其他文献
Sharonda LeBlanc的其他文献
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{{ truncateString('Sharonda LeBlanc', 18)}}的其他基金
Elucidating Molecular Mechanisms of Cancer Development by investigating Key DNA Repair Pathways
通过研究关键 DNA 修复途径阐明癌症发展的分子机制
- 批准号:
9756350 - 财政年份:2017
- 资助金额:
$ 13.16万 - 项目类别:
Elucidating molecular interactions of MutL in mismatch repair using single molecule FRET
使用单分子 FRET 阐明错配修复中 MutL 的分子相互作用
- 批准号:
9327441 - 财政年份:2017
- 资助金额:
$ 13.16万 - 项目类别:
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