Phosphorylation as a site-specific regulatory mechanism of the liquid-liquid phase separation of TDP-43 low complexity domain.
磷酸化作为 TDP-43 低复杂性结构域液-液相分离的位点特异性调节机制。
基本信息
- 批准号:10401259
- 负责人:
- 金额:$ 5.13万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-05-01 至 2024-04-30
- 项目状态:已结题
- 来源:
- 关键词:AffectAgingAlzheimer like pathologyAlzheimer&aposs DiseaseAmino AcidsAmyloidAmyotrophic Lateral SclerosisAntibodiesArginineAsparagineAspartic AcidBehaviorBrainC-terminalCellular Stress ResponseChargeComplexCoupledCytoplasmic GranulesDataDepositionDetectionDevelopmentDiseaseElectron Spin Resonance SpectroscopyElectrostaticsFluorescence Recovery After PhotobleachingFluorescence Resonance Energy TransferFrontotemporal Lobar DegenerationsGelImmobilizationIn VitroKnowledgeLabelLearningLiquid substanceMediatingMessenger RNAMethodsMicroscopyModelingModificationMolecularMonitorMutateMutationN-terminalNMR SpectroscopyNerve DegenerationNeurodegenerative DisordersNuclear Magnetic ResonanceOrganellesPathologicPathologyPatientsPhasePhosphorylationPhosphorylation SitePhosphotransferasesPhysiologicalPhysiological ProcessesPlayPost-Translational Protein ProcessingProcessPropertyProteinsRNARNA-Binding ProteinsResolutionRoleSamplingSchemeSepharoseSerineSignal TransductionSiteSodium ChlorideSpin LabelsStressStructureTechniquesTestingTherapeuticTimeTranslationsVariantWestern Blottingage relatedage related neurodegenerationbaseexpectationexperimental studyfluorophorefrontotemporal lobar dementia-amyotrophic lateral sclerosisgel electrophoresisin vivoinsightintermolecular interactionlight scatteringmonomernucleic acid binding proteinprotein TDP-43self assemblystress granulevirtual
项目摘要
Project Summary
The TAR DNA-binding protein of 43 kDa (TDP-43) is a nucleic-acid binding protein whose fragments have
been detected in pathological amyloid inclusions in the brains of patients with age-related neurodegenerative
disorders, including Alzheimer’s disease, amyotrophic lateral sclerosis, and frontotemporal lobar degeneration.
These fragments contain a region of TDP-43 called the low complexity domain (LCD), which has low amino acid
diversity and a propensity to form amyloid aggregates. The LCD also drives TDP-43 liquid-liquid phase
separation (LLPS), a phenomenon in which the protein self-associates into a reversible, dynamic, droplet-like
phase. LLPS is a physiologic process that mediates several functions of TDP-43, including its involvement in the
cellular stress response, but prolonged LLPS has been shown to promote pathological aggregation,
underscoring the need to understand how this process is regulated in vivo.
Phosphorylation of TDP-43 LCD has been well-documented in Alzheimer’s disease and other age-related
neurodegenerative disorders. Though the role of this modification remains largely unexplored for TDP-43, it does
influence LLPS in similar RNA-binding proteins such as FUS. To determine if LCD phosphorylation can regulate
TDP-43 LLPS, phosphomimetic variants of the LCD with Ser->Asp substitutions at known phosphorylation sites
were prepared. Phase separation of variants was studied using light-scattering and microscopy, and it was found
the phosphomimetic substitutions dramatically modulated LLPS in a site-specific manner. LCD variants with
phosphomimetic substitutions in the α-helical region, a part of the protein involved in protein-protein contacts,
displayed reduced LLPS-propensity; conversely, LCD variants with phosphomimetic substitutions in the C-
terminal region (CTR), which is known to be phosphorylated in disease, displayed enhanced LLPS-propensity.
To elucidate the mechanisms behind this site-specific modulation of LLPS, we propose a three-pronged
approach using nuclear magnetic resonance (NMR) spectroscopy, electron paramagnetic resonance (EPR)
spectroscopy and fluorescence resonance energy transfer (FRET). NMR will be employed to determine local
secondary structure of the phosphomimetic α-helical variant. As the α-helical region is known to be altered by
oligomerization, EPR will be used concurrently to see if the α-helical phosphomimetic substitution changes the
oligomerization behavior of this variant. Preliminary data suggest the CTR variants have higher LLPS-
propensities because of an intermolecular electrostatic interaction created by the phosphomimetic substitution
at the CTR. Our second aim will thus use FRET to verify the presence of this hypothesized intermolecular
interaction and probe which residues contribute to it. Furthermore, fluorescence recovery after photobleaching
(FRAP) will be employed to determine how these phosphomimetic substitutions alter the material properties of
TDP-43 LCD droplets. These approaches will provide mechanistic insights into how phosphorylation may site-
specifically direct TDP-43 LLPS and contribute to neurodegenerative pathologies like Alzheimer’s disease.
项目总结
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 }}
Raza Haider其他文献
Raza Haider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('Raza Haider', 18)}}的其他基金
Phosphorylation as a site-specific regulatory mechanism of the liquid-liquid phase separation of TDP-43 low complexity domain.
磷酸化作为 TDP-43 低复杂性结构域液-液相分离的位点特异性调节机制。
- 批准号:
10620157 - 财政年份:2021
- 资助金额:
$ 5.13万 - 项目类别:
Phosphorylation as a site-specific regulatory mechanism of the liquid-liquid phase separation of TDP-43 low complexity domain.
磷酸化作为 TDP-43 低复杂性结构域液-液相分离的位点特异性调节机制。
- 批准号:
10156401 - 财政年份:2021
- 资助金额:
$ 5.13万 - 项目类别:
相似海外基金
Interplay between Aging and Tubulin Posttranslational Modifications
衰老与微管蛋白翻译后修饰之间的相互作用
- 批准号:
24K18114 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
The Canadian Brain Health and Cognitive Impairment in Aging Knowledge Mobilization Hub: Sharing Stories of Research
加拿大大脑健康和老龄化认知障碍知识动员中心:分享研究故事
- 批准号:
498288 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Operating Grants
EMNANDI: Advanced Characterisation and Aging of Compostable Bioplastics for Automotive Applications
EMNANDI:汽车应用可堆肥生物塑料的高级表征和老化
- 批准号:
10089306 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Collaborative R&D
Baycrest Academy for Research and Education Summer Program in Aging (SPA): Strengthening research competencies, cultivating empathy, building interprofessional networks and skills, and fostering innovation among the next generation of healthcare workers t
Baycrest Academy for Research and Education Summer Program in Aging (SPA):加强研究能力,培养同理心,建立跨专业网络和技能,并促进下一代医疗保健工作者的创新
- 批准号:
498310 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Operating Grants
関節リウマチ患者のSuccessful Agingに向けたフレイル予防対策の構築
类风湿性关节炎患者成功老龄化的衰弱预防措施的建立
- 批准号:
23K20339 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Life course pathways in healthy aging and wellbeing
健康老龄化和福祉的生命历程路径
- 批准号:
2740736 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Studentship
NSF PRFB FY 2023: Connecting physiological and cellular aging to individual quality in a long-lived free-living mammal.
NSF PRFB 2023 财年:将生理和细胞衰老与长寿自由生活哺乳动物的个体质量联系起来。
- 批准号:
2305890 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Fellowship Award
I-Corps: Aging in Place with Artificial Intelligence-Powered Augmented Reality
I-Corps:利用人工智能驱动的增强现实实现原地老龄化
- 批准号:
2406592 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Standard Grant
McGill-MOBILHUB: Mobilization Hub for Knowledge, Education, and Artificial Intelligence/Deep Learning on Brain Health and Cognitive Impairment in Aging.
McGill-MOBILHUB:脑健康和衰老认知障碍的知识、教育和人工智能/深度学习动员中心。
- 批准号:
498278 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Operating Grants
Welfare Enhancing Fiscal and Monetary Policies for Aging Societies
促进老龄化社会福利的财政和货币政策
- 批准号:
24K04938 - 财政年份:2024
- 资助金额:
$ 5.13万 - 项目类别:
Grant-in-Aid for Scientific Research (C)