Imaging of Single HIV-1 Uncoating and Transport to the nucleus
Imaging of Single HIV-1 Uncoating and Transport to the nucleus
批准号:
9354023
负责人:
Gregory B Melikian
金额:
$59.65万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2017
资助国家:
美国
项目状态:
已结题
起止时间:
2017-03-24 至 2022-02-28
关键词:
AffectAffinityAgreementAvidityBindingBiochemicalBiological AssayCapsidCapsid ProteinsCell LineCell NucleusCell fusionCellsComplexConeCore ProteinCryoelectron MicroscopyCyclophilin ACytoplasmDNA IntegrationDNA biosynthesisDsRedEnzymesEventGeneticGenomeGoalsHIV-1Hela CellsHeterogeneityHumanImageImageryImaging DeviceIndividualInfectionInfection preventionIntegraseIntegration Host FactorsKinesinKineticsKnock-outLabelLengthLifeLinkMediatingMembraneMinorMutationNatureNuclear ImportNuclear PoreNuclear Pore ComplexNucleocapsidPhenotypeProcessProteinsPublishingRNA-Directed DNA PolymeraseRegulationReverse TranscriptionReverse Transcription InhibitionRibonuclease HSeriesStem cellsStructureTestingTubeViralViral GenomeVirionVirusVirus Integrationanterograde transportbasedesignflexibilitygenomic RNAimprovedinsightmacrophagemembermutantnovelnovel strategiesparticleprematurepreventspatiotemporalviral DNA
中文摘要
在病毒-细胞融合后,锥形HIV-1衣壳的降解是
造成终身感染HIV-1进入的这一步,被称为脱膜,是至关重要的,
不太了解。我们最近开发了一种新的策略来可视化HIV-1的脱壳,
是基于荧光标记的寡聚形式的captain结合宿主蛋白亲环素A
(CypA-DsRed)。CypA-DsRed通过高亲合力特异性包装成病毒粒子,
与HIV-1衣壳结合,不损害感染性。这个探测器仍然与
在病毒-细胞融合后具有核心,并且在脱壳时释放。支持这一观点的是我们的
发现来自单个融合后核心的CypA-DsRed损失率受到以下因素的调节:
影响核心稳定性的突变,并通过逆转录加速。CypA-DsRed
基于成像分析揭示了HIV-1脱壳的双相动力学,具有大量核心
融合后不久脱落衣壳蛋白。该试验还显示,
HeLa衍生细胞和原代人巨噬细胞中的未包被表型。我们提出
目的:(1)通过功能和结构描述CypA-DsRed与HIV-1核心的相互作用
CypA-DsRed/衣壳复合物的研究;(2)阐明了逆转录病毒与细胞凋亡之间的关系。
转录和单核脱壳;(3)研究宿主对HIV-1脱壳的调控
Nup 358和CPSF 6等因素。这些研究采用了基因、
生物化学和先进的成像工具,预计将提供关键的见解,
HIV-1在活细胞中脱壳的动力学和调节。
英文摘要
Disassembly of the cone-shaped HIV-1 capsid after virus-cell fusion is a prerequisite for
establishing a life-long infection. This step in HIV-1 entry, referred to as uncoating, is critical yet
poorly understood. We have recently developed a novel strategy to visualize HIV-1 uncoating that
is based on a fluorescently tagged oligomeric form of a capsid-binding host protein cyclophilin A
(CypA-DsRed). CypA-DsRed, which is specifically packaged into virions through the high-avidity
binding to the HIV-1 capsid, does not compromise the infectivity. This probe remains associated
with cores after virus-cell fusion and is released upon uncoating. Supporting this notion is our
finding that the rate of CypA-DsRed loss from individual post-fusion cores is modulated by
mutations affecting the core stability and is accelerated by reverse transcription. The CypA-DsRed
based imaging assay revealed a biphasic kinetic of HIV-1 uncoating, with a large number of cores
shedding the capsid protein shortly after fusion. This assay also revealed marked differences in
the uncoating phenotype in HeLa-derived cells and primary human macrophages. We propose
to: (1) delineate CypA-DsRed interactions with HIV-1 core through functional and structural
studies of CypA-DsRed/capsid complexes; (2) elucidate the relationship between reverse
transcription and single core uncoating; and (3) investigate regulation of HIV-1 uncoating by host
factors, such as Nup358 and CPSF6. These studies, employing a combination of genetic,
biochemical and advanced imaging tools, are expected to provide critical insights into the
dynamics and regulation of HIV-1 uncoating in living cells.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Biophysics Core
-
批准号:10508448
-
项目类别:
-
资助金额:$77.98万
-
财政年份:2022
-
负责人:Gregory B Melikian
-
依托单位:
Biophysics Core
-
批准号:10650878
-
项目类别:
-
资助金额:$80.21万
-
财政年份:2022
-
负责人:Gregory B Melikian
-
依托单位:
Molecular Interactions of HIV-1 with the Nuclear Pore Complex
-
批准号:10241258
-
项目类别:
-
资助金额:$136.92万
-
财政年份:2019
-
负责人:Gregory B Melikian
-
依托单位:
Molecular Interactions of HIV-1 with the Nuclear Pore Complex
-
批准号:10462620
-
项目类别:
-
资助金额:$134.88万
-
财政年份:2019
-
负责人:Gregory B Melikian
-
依托单位:
Inhibition of viral entry by interferon-induced proteins
-
批准号:10418696
-
项目类别:
-
资助金额:$35.6万
-
财政年份:2018
-
负责人:Gregory B Melikian
-
依托单位:
Inhibition of viral entry by interferon-induced proteins
-
批准号:10190798
-
项目类别:
-
资助金额:$35.6万
-
财政年份:2018
-
负责人:Gregory B Melikian
-
依托单位:
Kinetic Determinants of HIV Neutralization
-
批准号:7929311
-
项目类别:
-
资助金额:$23.25万
-
财政年份:2010
-
负责人:Gregory B Melikian
-
依托单位:
Kinetic Determinants of HIV Neutralization
-
批准号:8142878
-
项目类别:
-
资助金额:$19.18万
-
财政年份:2010
-
负责人:Gregory B Melikian
-
依托单位:
Functional Characterization of the Hepatitis C Virus E1-E2 Glycoproteins
-
批准号:7522862
-
项目类别:
-
资助金额:$18.75万
-
财政年份:2009
-
负责人:Gregory B Melikian
-
依托单位:
Functional Characterization of the Hepatitis C Virus E1-E2 Glycoproteins
-
批准号:8116923
-
项目类别:
-
资助金额:$15.55万
-
财政年份:2009
-
负责人:Gregory B Melikian
-
依托单位:
Functional Characterization of the Hepatitis C Virus E1-E2 Glycoproteins
-
批准号:7897844
-
项目类别:
-
资助金额:$3.7万
-
财政年份:2009
-
负责人:Gregory B Melikian
-
依托单位:
The entry mechanism used by a model retrovirus.
-
批准号:7008544
-
项目类别:
-
资助金额:$35.76万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
Entry Mechanisms used by a model retrovirus
-
批准号:7370296
-
项目类别:
-
资助金额:$36.25万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
The Mechanism of Arenavirus Entry into Cells
-
批准号:10623143
-
项目类别:
-
资助金额:$41.76万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
Entry mechanisms used by a model retrovirus
-
批准号:8577613
-
项目类别:
-
资助金额:$36.66万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
Entry mechanisms used by a model retrovirus
-
批准号:8656664
-
项目类别:
-
资助金额:$36.66万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
The Mechanism of Arenavirus Entry into Cells
-
批准号:9889875
-
项目类别:
-
资助金额:$41.76万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
Entry Mechanisms used by a model retrovirus
-
批准号:8116813
-
项目类别:
-
资助金额:$38.29万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
The Mechanism of Arenavirus Entry into Cells
-
批准号:9755767
-
项目类别:
-
资助金额:$44.56万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
The entry mechanism used by a model retrovirus.
-
批准号:7176092
-
项目类别:
-
资助金额:$35.71万
-
财政年份:2003
-
负责人:Gregory B Melikian
-
依托单位:
海外基金