Direct Visualization of HIV-1 with Correlative Live-Cell Microscopy and Cryo-Electron Tomography

被引：72

作者：

Jun, Sangmi ^{[1
]}

Ke, Danxia ^{[1
]}

Debiec, Karl ^{[1
]}

Zhao, Gongpu ^{[1
]}

Meng, Xin ^{[1
]}

Ambrose, Zandrea ^{[2
]}

Gibson, Gregory A. ^{[3
]}

Watkins, Simon C. ^{[3
]}

Zhang, Peijun ^{[1
]}

机构：

[1] Univ Pittsburgh, Sch Med, Dept Biol Struct, Pittsburgh, PA 15260 USA

[2] Univ Pittsburgh, Sch Med, Div Infect Dis, Dept Med, Pittsburgh, PA 15260 USA

[3] Univ Pittsburgh, Sch Med, Dept Cell Biol & Physiol, Pittsburgh, PA 15260 USA

来源：

STRUCTURE | 2011年 / 19卷 / 11期

基金：

美国国家卫生研究院;

关键词：

HERPES-SIMPLEX-VIRUS; ELECTRON CRYOTOMOGRAPHY; STRUCTURAL ORGANIZATION; REVERSE TRANSCRIPTION; LIGHT-MICROSCOPY; RECEPTOR ARRAYS; VACCINIA VIRUS; LIVING CELLS; VIRIONS; FUSION;

D O I：

10.1016/j.str.2011.09.006

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cryo-electron tomography (cryoET) allows 3D visualization of cellular structures at molecular resolution in a close-to-native state and therefore has the potential to help elucidate early events of HIV-1 infection in host cells. However, structural details of infecting HIV-1 have not been observed, due to technological challenges in working with rare and dynamic HIV-1 particles in human cells. Here, we report structural analysis of HIV-1 and host-cell interactions by means of a correlative high-speed 3D live-cell-imaging and cryoET method. Using this method, we showed under near-native conditions that intact hyperstable mutant HIV-1 cores are released into the cytoplasm of host cells. We further obtained direct evidence to suggest that a hyperstable mutant capsid, E45A, showed delayed capsid disassembly compared to the wild-type capsid. Together, these results demonstrate the advantages of our correlative live-cell and cryoET approach for imaging dynamic processes, such as viral infection.

引用

页码：1573 / 1581

页数：9

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