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

作者：Jun Sangmi; Ke Danxia; Debiec Karl; Zhao Gongpu; Meng Xin; Ambrose Zandrea; Gibson Gregory A; Watkins Simon C; Zhang Peijun^*

来源：Structure, 2011, 19(11): 1573-1581.

DOI：10.1016/j.str.2011.09.006

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.

• 出版日期2011-11-9