Regulation of unwinding and remodeling activities in FeS-DNA helicases
FeS-DNA 解旋酶解旋和重塑活性的调节
基本信息
- 批准号:9022495
- 负责人:
- 金额:$ 28万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-06-01 至 2019-02-28
- 项目状态:已结题
- 来源:
- 关键词:ATP phosphohydrolaseAffectAgingAntineoplastic AgentsBRCA1 geneBindingBinding SitesBiochemicalCell physiologyChemistryComplexDNADNA BindingDNA DamageDNA MaintenanceDNA RepairDNA StructureDNA lesionDataDefectDiagnosisDiseaseDrug TargetingEnzymesExogenous FactorsFamilyFanconi&aposs AnemiaFluorescence MicroscopyFoundationsFrequenciesG-QuartetsGenerationsGenetic RecombinationGenetic TranscriptionGenomeGenomic InstabilityHDAC1 geneHealthHereditary DiseaseHuman ActivitiesHypertensionIndividualIronLabelLearningLifeLinkMaintenanceMalignant NeoplasmsMediatingMethodologyMismatch RepairMolecular ConformationMolecular MotorsMotionMotorMutationNucleoproteinsNucleotide Excision RepairPositioning AttributePredispositionProcessProgeriaProteinsRegulationRoleSiteSulfurSymptomsTestingTimeTrichothiodystrophyTumor Suppressor Proteinsage relateddesigngenome integrityhelicasehuman diseaseinhibitor/antagonistinnovationmalignant breast neoplasmmutantnovelprotein protein interactionreconstitutionsingle moleculesynthetic constructtargeted treatmenttranslocase
项目摘要
DESCRIPTION (provided by applicant): This application focuses on iron-sulfur containing (FeS) helicases, a prominent DNA helicase family whose deficiency or dysregulation is linked to human diseases ranging from cancer predisposition to hypertension. In addition to the Superfamily II motor core, FeS helicases possess two family specific auxiliary domains: an FeS cluster domain and an ARCH domain. The secondary DNA binding site formed with the help of the auxiliary domains which positions the helicase in an orientation to unwind duplex, controls the helicase rate, and verifies the integrity of the translocating strand. I propose that the frequency of ARCH domain opening and closing in FeS helicases modulates their activities. We will use this helicase family to test for the first time how the exogenous factors affect the mechano-chemistry of the helicases through modulating the frequency of its core and auxiliary domains motions. Our objective is to determine the mechanism by which the domain mobility controls the activities of three FeS helicases, XPD, FANCJ and RTEL1. To achieve this objective we will use a synergistic set of biochemical reconstitutions and novel single-molecule methodologies developed in my lab. Aim 1: Determine the role of ARCH domain mobility in controlling XPD activities. We will build on our preliminary data showing that the cognate DNA lesions stabilize the closed conformation of the ARCH. Using single-molecule total internal reflection fluorescence microscopy (TIRFM), we will observe domain motions of individual fluorescently labeled XPD molecules as they interact with DNA. We will learn how ARCH domain motions control activities of XPD helicase and its malfunction in disease. Aim 2: Determine the role of ARCH domain mobility in FANCJ and RTEL1 mediated DNA unwinding and remodeling of G-quadruplexes. Upon completion of this aim we will learn how the helicase and G-quadruplex remodeling activities of FANCJ and RTEL1 correlate with ARCH domain motions. We will also learn how FANCJ mutations associated with breast cancer and Fanconi Anemia perturb FANCJ activities, ARCH domain mobility and the ability to discriminate between damaged and damage-free DNA. Aim 3: Determine how protein partners tune the activities of FANCJ and RTEL1. We will test the hypothesis that interactions with key protein partners (BRCA1 tumor suppressor protein, hMLH1 mismatch repair protein and PCNA clamp) govern helicase and translocase activities by modifying domain mobility of FANCJ and RTEL1. Together, the anticipated results of the three proposed aims will not only close the gaps in the mechanistic understanding of how helicases' distinct biochemical activities are regulated, but also identify explicit strategies to selectively modulate them. This information will pave the way for the design of inhibitors of FANCJ or RTEL1 to be used to target specific aspects of cancer and aging related diseases.
描述(申请人提供):本申请重点研究含铁硫解旋酶(FES)解旋酶,这是一个重要的DNA解旋酶家族,其缺乏或失调与从癌症易感性到高血压等人类疾病有关。除了超家族II运动核心外,FES解旋酶还具有两个家族特有的辅助结构域:FES簇区和ARCH结构域。在辅助结构域的帮助下形成的二级DNA结合位点将解旋酶定位在一个方向以解开双链,控制解旋酶的速率,并验证转移链的完整性。我认为FES解旋酶的ARCH结构域的开启和关闭的频率调节了它们的活性。我们将利用这个解旋酶家族首次测试外源因素如何通过调节其核心和辅助结构域的运动频率来影响解旋酶的机械力化学。我们的目标是确定结构域迁移率控制三种FES解旋酶XPD、FANCJ和RTEL1活性的机制。为了实现这一目标,我们将使用一套协同的生化重组和在我的实验室开发的新颖的单分子方法。目的1:确定ARCH结构域迁移率在控制XPD活性中的作用。我们将以我们的初步数据为基础,表明同源DNA损伤稳定了拱门的闭合构象。使用单分子全内反射荧光显微镜(TIRFM),我们将观察单个荧光标记的XPD分子与DNA相互作用时的结构域运动。我们将学习ARCH结构域运动如何控制XPD解旋酶的活性及其在疾病中的功能障碍。目的:探讨ARCH结构域迁移率在FANCJ和RTEL1介导的G-四链DNA解离和重塑中的作用。在完成这一目标后,我们将了解FANCJ和RTEL1的解旋酶和G-四链重塑活动如何与ARCH结构域运动相关。我们还将了解与乳腺癌和Fanconi贫血相关的FANCJ突变如何扰乱FANCJ的活性、ARCH结构域的流动性以及区分受损和无损伤DNA的能力。目的3:确定蛋白质伙伴如何调节FANCJ和RTEL1的活性。我们将检验这一假设,即与关键蛋白质伙伴(BRCA1肿瘤抑制蛋白、hMLH1错配修复蛋白和增殖细胞核抗原钳)的相互作用通过改变FANCJ和RTEL1的结构域迁移率来控制解旋酶和易位酶的活性。总之,这三个拟议目标的预期结果不仅将弥合对解旋酶不同生化活动如何调节的机械性理解方面的差距,而且还将确定有选择地调节它们的明确策略。这些信息将为FANCJ或RTEL1抑制剂的设计铺平道路,以用于针对癌症和衰老相关疾病的特定方面。
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 }}
Maria Spies其他文献
Maria Spies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('Maria Spies', 18)}}的其他基金
Natural products inhibitors targeting homology-directed DNA repair for cancer therapy
针对癌症治疗的同源定向 DNA 修复的天然产物抑制剂
- 批准号:
10651048 - 财政年份:2023
- 资助金额:
$ 28万 - 项目类别:
Lumick's C-Trap instrument for single-molecule analysis of macromolecular dynamics
Lumick 用于大分子动力学单分子分析的 C-Trap 仪器
- 批准号:
10175508 - 财政年份:2021
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
10808780 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
10377656 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
10593161 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
9900829 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
10375412 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Assembly and Dynamics of Molecular Machines in Genome Maintenance
基因组维护中分子机器的组装和动力学
- 批准号:
10798482 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
FASEB SRC on Helicases and Nucleic-Acid Based Machines: From Mechanism to Insights into Disease
FASEB SRC 关于解旋酶和核酸机器:从机制到洞察疾病
- 批准号:
8986287 - 财政年份:2015
- 资助金额:
$ 28万 - 项目类别:
Regulation of unwinding and remodeling activities in FeS-DNA helicases
FeS-DNA 解旋酶解旋和重塑活性的调节
- 批准号:
9222028 - 财政年份:2014
- 资助金额:
$ 28万 - 项目类别:
相似海外基金
Hormone therapy, age of menopause, previous parity, and APOE genotype affect cognition in aging humans.
激素治疗、绝经年龄、既往产次和 APOE 基因型会影响老年人的认知。
- 批准号:
495182 - 财政年份:2023
- 资助金额:
$ 28万 - 项目类别:
Parkinson's disease and aging affect neural activation during continuous gait alterations to the split-belt treadmill: An [18F] FDG PET Study.
帕金森病和衰老会影响分体带跑步机连续步态改变期间的神经激活:[18F] FDG PET 研究。
- 批准号:
400097 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
The elucidation of the mechanism by which intestinal epithelial cells affect impaired glucose tolerance during aging
阐明衰老过程中肠上皮细胞影响糖耐量受损的机制
- 批准号:
19K09017 - 财政年份:2019
- 资助金额:
$ 28万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Does aging of osteocytes adversely affect bone metabolism?
骨细胞老化会对骨代谢产生不利影响吗?
- 批准号:
18K09531 - 财政年份:2018
- 资助金额:
$ 28万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Links between affect, executive function, and prefrontal structure in aging: A longitudinal analysis
衰老过程中情感、执行功能和前额叶结构之间的联系:纵向分析
- 批准号:
9766994 - 财政年份:2018
- 资助金额:
$ 28万 - 项目类别:
Affect regulation and Beta Amyloid: Maturational Factors in Aging and Age-Related Pathology
影响调节和 β 淀粉样蛋白:衰老和年龄相关病理学中的成熟因素
- 批准号:
9320090 - 财政年份:2017
- 资助金额:
$ 28万 - 项目类别:
Affect regulation and Beta Amyloid: Maturational Factors in Aging and Age-Related Pathology
影响调节和 β 淀粉样蛋白:衰老和年龄相关病理学中的成熟因素
- 批准号:
10166936 - 财政年份:2017
- 资助金额:
$ 28万 - 项目类别:
Affect regulation and Beta Amyloid: Maturational Factors in Aging and Age-Related Pathology
影响调节和 β 淀粉样蛋白:衰老和年龄相关病理学中的成熟因素
- 批准号:
9761593 - 财政年份:2017
- 资助金额:
$ 28万 - 项目类别:
Experimental Model of Depression in Aging: Insomnia, Inflammation, and Affect Mechanisms
衰老过程中抑郁症的实验模型:失眠、炎症和影响机制
- 批准号:
9925164 - 财政年份:2016
- 资助金额:
$ 28万 - 项目类别:
Experimental Model of Depression in Aging: Insomnia, Inflammation, and Affect Mechanisms
衰老过程中抑郁症的实验模型:失眠、炎症和影响机制
- 批准号:
9345997 - 财政年份:2016
- 资助金额:
$ 28万 - 项目类别: