Quantitative microscopy of functional HIV post-entry complexes reveals association of replication with the viral capsid

• DOI: 10.7554/elife.04114
• Publication Date: 2014-12-17T12:34:32Z
• Authors: Ke Peng; Walter Muranyi; Bärbel Glass; Vibor Laketa; Stephen R Yant; Luong Tsai; Tomas Cihlar; Barbara Müller; Hans-Georg Kräuss...
• Keywords: Transcription; Host factors

The steps from HIV-1 cytoplasmic entry until integration of the reverse transcribed genome are currently enigmatic. They occur in ill-defined reverse-transcription- and pre-integration-complexes (RTC, PIC) with various host viral proteins implicated. In this study, we report quantitative detection functional RTC/PIC by labeling nascent DNA combined integrase. We show that CA (capsid) protein remains associated but is lost on nuclear PIC a HeLa-derived cell line. contrast, were almost always CA-positive primary human macrophages, indicating import capsids or capsid-like structures. further CA-targeted inhibitor PF74 exhibits bimodal mechanism, blocking association factor CPSF6 at low, abrogating transcription high concentrations. newly developed system ideally suited for studying retroviral post-entry events roles factors including sensors signaling molecules.