Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function.
微管蛋白二聚体调节途径的机制及其对微管功能的影响。
基本信息
- 批准号:10212401
- 负责人:
- 金额:$ 33.76万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2015
- 资助国家:美国
- 起止时间:2015-01-01 至 2024-05-31
- 项目状态:已结题
- 来源:
- 关键词:AddressBindingBiochemicalBiogenesisBiological AssayBiophysicsCatalysisCell divisionCellsChromosome SegregationComplexCryoelectron MicroscopyCytoplasmCytoskeletonDataDefectDevelopmentDimerizationDockingElementsEncephalopathiesEnzymesEukaryotic CellFission YeastFluorescence MicroscopyFundingGTP-Binding ProteinsGenesGrowthGuanosine TriphosphateGuanosine Triphosphate PhosphohydrolasesHereditary DiseaseHeterodimerizationHumanImageImage AnalysisImpairmentIn VitroIndividualInheritedKenny-Caffey syndromeLeadLife Cycle StagesLinkMalignant NeoplasmsMediatingMethodsMicrotubule PolymerizationMicrotubulesModelingMolecularMolecular ConformationMutationOrthologous GenePathway interactionsPatientsPhysiologicalPlus End of the MicrotubulePolymerasePolymersProcessPropertyProteinsPublic HealthPublishingQuality ControlReagentRegulationRegulatory PathwayResolutionRoleShapesSourceStructureSystemTestingTubulinWorkYeastsalpha Tubulinbasebeta Tubulincofactordesigndevelopmental diseasedimergiant axonal neuropathyimaging approachin vitro Assayin vivoin vivo imaginglissencephalylive cell imagingmutantneoplastic cellnervous system disorderoverexpressionparticlepolymerizationreconstitutionrecruitself assemblystemtooltraffickingtumortumor growth
项目摘要
Project Summary
The dynamic microtubule cytoskeleton mediates intracellular organization, generates forces in dividing or migrating
eukaryotic cells, and forms tracks for intracellular trafficking. The fundamental properties of microtubules, including
polarized growth and “dynamic instability”, stem directly from the activities of the microtubule building blocks, the α- and
β-tubulin heterodimers. Three conserved tubulin cofactors and dedicated Arf-like 2 G-protein form multi-subunit platforms
for the biogenesis and degradation of αβ-tubulin dimer, leading to a high concentration within the cytoplasm. The
mechanisms for these assemblies remain mostly mysterious, due in part to a lack of structural information. In addition, we
do not understand how conserved microtubule polymerases with arrays of Tumor Overexpressed Gene (TOG) domains
recruit αβ-tubulins and accelerate their incorporation while tracking dynamic microtubule ends. Understanding these
cellular pathways is critical since genetic defects that impair either soluble αβ-tubulin biogenesis or microtubule
polymerases are linked to inherited neurological and developmental disorders and are observed in human cancers,
respectively. This proposal explores the biochemical and physical mechanisms of αβ-tubulin biogenesis and microtubule
polymerase assemblies and their impact on microtubule function. Our strategy combines methods across multiple resolution
scales, including in vitro reconstitution of purified protein assemblies, structural studies by cryo-electron microscopy (cryo-
EM), reconstitution of assemblies with microtubule dynamics using in vitro fluorescence microscopy-based assays, and in
vivo live imaging with microtubules within living cells.
First, we will determine structural transitions describing αβ-tubulin biogenesis assemblies and their functional impact of
αβ-tubulin biogenesis and degradation. During the previous period, we established reconstitution system for these
assemblies with αβ-tubulin and describe cryo-EM structural studies leading to medium resolution structures in complex
with αβ-tubulins. 1) We will determine structural states for the αβ-tubulin biogenesis assemblies in multiple biochemical
states using high-resolution cryo-EM to understand how these assemblies catalyze dimerization of αβ-tubulin and its
degradation. 2) We will dissect functional roles of structural elements and interactions within current structures to determine
their role in the αβ-tubulin biogenesis process using in vitro and in vivo methods. Second, we will examine the mechanisms
of microtubule polymerases with arrays of TOG domains their regulatory mechanisms. In the previous period, we describe
a new model for αβ-tubulin recruitment and polymerization by TOG domain arrays as microtubule polymerases, developed
based on our structural and biochemical studies. We validated this model using in vitro reconstitution and in vivo live
imaging of structure-based designer defective mutants, revealing that the αβ-tubulin accelerating and processive plus-end
tracking activities originate from unique features in TOG domain arrays. 1) We will study mechanisms of super-complexes
of microtubule polymerase in complex with their activators, the transforming acidic coiled-coil proteins, in by using well-
explored structural, in vitro reconstitution and in vivo live imaging strategies. 2) Determine the structural and functional
relevance of our new model to mammalian microtubule polymerases with their unique pentameric TOG domain array
arrangement using cryo-EM structural studies, in vitro reconstitution of designer mutants, and in vivo imaging approaches
of structure-based mutants. We expect these studies to yield new structural and biophysical data, which will refine our new
models will deepen our understanding of soluble αβ-tubulin biogenesis, recruitment and incorporation during microtubule
polymerization. This understanding will in turn point toward new strategies for addressing defects in tubulin biogenesis and
regulation, potentially impacting patients with a range of developmental and neurological disorders.
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)
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Jawdat MH Al-Bassam的其他文献
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{{ truncateString('Jawdat MH Al-Bassam', 18)}}的其他基金
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function
微管蛋白二聚体调控途径的机制及其对微管功能的影响
- 批准号:
10219718 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function.
微管蛋白二聚体调节途径的机制及其对微管功能的影响。
- 批准号:
10053131 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function.
微管蛋白二聚体调节途径的机制及其对微管功能的影响。
- 批准号:
10414979 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin Dimer Regulatory Pathways and Their Impact on Microtubule Function
微管蛋白二聚体调控途径的机制及其对微管功能的影响
- 批准号:
10625195 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function
微管蛋白二聚体调控途径的机制及其对微管功能的影响
- 批准号:
8818688 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function
微管蛋白二聚体调控途径的机制及其对微管功能的影响
- 批准号:
10390213 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Mechanisms of Tubulin dimer Regulatory Pathways and their impact on Microtubule Function.
微管蛋白二聚体调节途径的机制及其对微管功能的影响。
- 批准号:
10619579 - 财政年份:2015
- 资助金额:
$ 33.76万 - 项目类别:
Microtubule Polymerization and Depolymerization Mechanisms by Conserved Proteins
保守蛋白的微管聚合和解聚机制
- 批准号:
8042154 - 财政年份:2010
- 资助金额:
$ 33.76万 - 项目类别:
Microtubule Polymerization and Depolymerization Mechanisms by Conserved Proteins
保守蛋白的微管聚合和解聚机制
- 批准号:
7450236 - 财政年份:2008
- 资助金额:
$ 33.76万 - 项目类别:
Microtubule Polymerization and Depolymerization Mechanisms by Conserved Proteins
保守蛋白的微管聚合和解聚机制
- 批准号:
8327133 - 财政年份:2008
- 资助金额:
$ 33.76万 - 项目类别:
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