Autophagosome closure by the ESCRT machinery
Autophagosome closure by the ESCRT machinery
批准号:
10453304
负责人:
HONG-GANG WANG
金额:
$34.81万
依托单位国家:
美国
项目类别:
财政年份:
2018
资助国家:
美国
项目状态:
未结题
起止时间:
2018-07-01 至 2026-07-31
关键词:
ATP phosphohydrolaseAddressAgingAutophagocytosisAutophagosomeBindingBiogenesisBiological AssayCRISPR/Cas technologyCell membraneCellular MembraneComplexCytokinesisDataDefectDiseaseElectron Transport Complex IIIElementsEndosomesEventExcisionFamily memberGenesGoalsGrowthHealthHeart DiseasesHomologous GeneHumanKnowledgeLibrariesLipidsLiver diseasesLysosomesMalignant NeoplasmsMammalian CellMediatingMembraneModelingMolecularN-terminalNerve DegenerationNuclear EnvelopePathologic ProcessesPhysiological ProcessesPositioning AttributeProcessProtein DatabasesProtein FamilyProteinsProteomicsRecyclingRoleSiteSorting - Cell MovementStressStructureTSG101 geneTertiary Protein StructureTestingTextUbiquitinUbiquitinationUrsidae FamilyVariantVesicleWorkbasegenome-widehuman diseasemembrane modelnovelprotein functionreceptorrecruitrepairedresponsesealspatiotemporal
中文摘要
项目摘要/摘要
这个项目的目标是解决关于吞噬细胞如何闭合形成的知识上的一个根本差距。
双膜自噬小体。在巨型自噬期间(下称自噬),呈新月形
吞噬体在细胞质周围伸长并封闭以产生双膜自噬小体
与溶酶体融合以促进货物降解。当噬菌体边缘变窄时,膜必须经历
在类似于内体分选的过程中分裂分离内膜和外膜
转运所需的复合体(ESCRT)介导的膜断裂。使用我们优雅的HaloTag-LC3
自噬完成分析,我们为ESCRT机制提供了第一个实验证据。
并确定ESCRT-I亚单位VPS37A是导致ESCRT-I亚基关闭的关键因素
将下游的ESCRT重新招募到噬菌体。值得注意的是,我们发现N末端推定
VPS37A的泛素E2变异体(PUEV)结构域是自噬小体关闭所必需的,但
对于ESCRT介导的其他膜脱落过程,包括内小体受体分选,是必不可少的
和胞质分裂。隔室特异性靶向因子启动了四种ESCRT的顺序招募
复合体(ESCRT-I、-II、-III和Vps4)到膜断裂部位。而吞噬动物特有的
我们的初步研究表明,含有VPS37A的ESCRT-I复合体的靶向因子尚不清楚
发现VPS37A PUEV与含有阴离子脂类和脂类的高度弯曲的膜相互作用
包装缺陷,这都是噬菌体边缘的特征。我们假设PUEV有选择地
与高度弯曲的吞噬体膜相互作用,将ESCRT-I靶向噬菌体。此外,我们的
初步工作表明,ESCRT在吞噬细胞中的募集需要蛋白质泛素化,并且
LC3/GABARAP接合机制,使我们相信稳定的膜缔合
ESCRT-I需要与与噬菌体相关的泛素化货物进行额外的相互作用。我们正处在一个理想中
在以下特定目标中测试我们的假设:(1)确定VPS37A如何靶向ESCRT-I
在自噬小体生物发生过程中吞噬并指导下游ESCRT的组装;(2)
确定自噬过程中ESCRT-I的噬菌体特异性靶向因子。因为自噬参与了
众多的生理和病理过程,这些研究将对
人类的健康和疾病。
英文摘要
Project Summary/Abstract
The goal of this project is to address a fundamental gap in knowledge on how phagophores are closed to form
double membrane autophagosomes. During macroautophagy (hereafter autophagy), crescent-shaped
phagophores elongate around cytoplasmic material and seal to generate double-membrane autophagosomes
that fuse with lysosomes for cargo degradation. As the phagophore rim narrows, the membranes must undergo
fission to separate the inner and outer membranes in a process that bears resemblance to endosomal sorting
complexes required for transport (ESCRT)-mediated membrane scission. Using our elegant HaloTag-LC3
autophagosome completion assay, we provided the first experimental evidence for the ESCRT machinery in
mammalian phagophore closure and identified the ESCRT-I subunit VPS37A as critical factor for the
recruitment of downstream ESCRTs to the phagophore. Notably, we found that the N-terminal putative
ubiquitin E2 variant (PUEV) domain of VPS37A is uniquely required for autophagosome closure but is
dispensable for other ESCRT-mediated membrane abscission processes, including endosome receptor sorting
and cytokinesis. Compartment-specific targeting factors initiate the sequential recruitment of the four ESCRT
complexes (ESCRT-I, -II, -III and VPS4) to the membrane scission site. While the phagophore-specific
targeting factors for the VPS37A-containing ESCRT-I complex are unknown, our preliminary study has
revealed that VPS37A PUEV interacts with highly curved membranes containing anionic lipids and lipid
packing defects, which are all features of the phagophore rim. We hypothesize that the PUEV selectively
interacts with highly curved phagophore membranes to target ESCRT-I to the phagophore. Furthermore, our
preliminary work has revealed that ESCRT recruitment to phagophores requires protein ubiquitylation and the
LC3/GABARAP conjugation machinery, leading us to believe that the stabilization of membrane-associated
ESCRT-I requires additional interactions with phagophore-associated ubiquitylated cargo. We are in an ideal
position to test our hypotheses in the following Specific Aims: (1) to determine how VPS37A targets ESCRT-I
to phagophores and directs the assembly of downstream ESCRTs during autophagosome biogenesis; (2) to
identify phagophore-specific targeting factors for ESCRT-I during autophagy. As autophagy is involved in
numerous physiological and pathological processes, these studies will have far-reaching implications for
human health and disease.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Autophagy Heterogeneity and Tumor Metastasis
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批准号:10212775
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项目类别:
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资助金额:$41.7万
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财政年份:2021
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负责人:HONG-GANG WANG
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依托单位:
Autophagosome closure by the ESCRT machinery
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批准号:10383918
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资助金额:$7.5万
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财政年份:2018
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依托单位:
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批准号:10448458
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资助金额:$34.14万
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依托单位:
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批准号:10132346
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资助金额:$31.22万
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负责人:HONG-GANG WANG
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依托单位:
Non-canonical Caspase-8 Activation on Autophagosomal Membranes
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批准号:10214562
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项目类别:
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资助金额:$34.85万
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财政年份:2018
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负责人:HONG-GANG WANG
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依托单位:
Non-canonical Caspase-8 Activation on Autophagosomal Membranes
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批准号:9983008
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项目类别:
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资助金额:$34.86万
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财政年份:2018
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依托单位:
Autophagosome closure by the ESCRT machinery
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批准号:9884792
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项目类别:
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资助金额:$31.23万
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财政年份:2018
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依托单位:
Autophagosome closure by the ESCRT machinery
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批准号:10703381
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资助金额:$34.8万
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依托单位:
Targeting autophagy in the TME
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批准号:8395589
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项目类别:
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资助金额:$16.64万
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财政年份:2012
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负责人:HONG-GANG WANG
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依托单位:
Targeting autophagy in the TME
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批准号:8495299
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项目类别:
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资助金额:$18.77万
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财政年份:2012
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负责人:HONG-GANG WANG
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依托单位:
Regulation of Autophagy and Tumorigenesis by Bif-1
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批准号:8016110
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项目类别:
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资助金额:$31.22万
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财政年份:2009
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负责人:HONG-GANG WANG
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依托单位:
Regulation of Autophagy and Tumorigenesis by Bif-1
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批准号:8447365
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项目类别:
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资助金额:$29.34万
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财政年份:2009
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负责人:HONG-GANG WANG
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依托单位:
Regulation of Autophagy and Tumorigenesis by Bif-1
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资助金额:$31.22万
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财政年份:2009
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负责人:HONG-GANG WANG
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依托单位:
Regulation of Autophagy and Tumorigenesis by Bif-1
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批准号:7653080
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项目类别:
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资助金额:$32.13万
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财政年份:2009
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负责人:HONG-GANG WANG
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依托单位:
Regulation of Autophagy and Tumorigenesis by Bif-1
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批准号:7798244
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项目类别:
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资助金额:$32.18万
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财政年份:2009
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负责人:HONG-GANG WANG
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依托单位:
Development of Small Molecule Antagonists of Bcl2/BclXL
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批准号:7214568
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财政年份:2006
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负责人:HONG-GANG WANG
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依托单位:
Mechanisms of Rad9 Mediated Checkpoints and Apoptosis
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批准号:6721374
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项目类别:
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资助金额:$24.11万
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财政年份:2002
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负责人:HONG-GANG WANG
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依托单位:
Mechanisms of Rad9 Mediated Checkpoints and Apoptosis
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批准号:6471623
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项目类别:
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资助金额:$24.11万
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财政年份:2002
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负责人:HONG-GANG WANG
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依托单位:
Mechanisms of Rad9 Mediated Checkpoints and Apoptosis
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批准号:6871234
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项目类别:
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资助金额:$24.11万
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财政年份:2002
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负责人:HONG-GANG WANG
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依托单位:
Mechanisms of Rad9 Mediated Checkpoints and Apoptosis
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批准号:7036487
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资助金额:$23.54万
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财政年份:2002
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负责人:HONG-GANG WANG
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依托单位:
海外基金