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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.

Original publication

DOI

10.1016/j.str.2011.09.006

Type

Journal article

Journal

Structure

Publication Date

09/11/2011

Volume

19

Pages

1573 - 1581

Keywords

Capsid Proteins, Cryoelectron Microscopy, Electron Microscope Tomography, HIV Infections, HIV-1, HeLa Cells, Host-Pathogen Interactions, Human Immunodeficiency Virus Proteins, Humans, Mutation, Missense, Single-Cell Analysis, Virion