Real-Time Imaging of Single HIV-1 Disassembly with Multicolor Viral Particles

Viral disassembly is poorly understood and related to the infection mechanism. However, directly observing the process in living cells remains technically challenging. In this study, the genome RNA, capsid, and matrix protein of the HIV-1 virus were labeled with a Ru(II) complex ([Ru(phen),(dppz)(2+)), the TC-FLAsH/ReAsH system, and EGFP/ECFP, respectively. Using the multi-colored virus and single-particle imaging, we were able to track the sequential disassembly process of single HIV-1 virus particles in live host cells. Approximately 0.1% of viral particles were observed to undergo a sequential disassembly process at 60-120 min post infection. The timing and efficiency of the disassembly were influenced by the cellular factor CypA and reverse transcription. The findings facilitate a better understanding of the processes governing the HIV-1 lifecycle. The multicolor labeling protocol developed in this study may find many applications involving virus host-cell interactions.