Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 5, Pages 2648-2654Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c05199
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Funding
- Strategic Priority Research Program of the Chinese Academy of Sciences, China [XDB29050100]
- National Natural Science Foundation [21890743]
- National Key Research and Development Program of China [2017YFA0205503]
- Youth Innovation Promotion Association CAS [2021359]
- Natural Science Foundation of Guangdong [2018B030306046]
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This study observed the sequential relationship between viral uncoating and reverse transcription by real-time imaging of single HIV-1 particles. It was found that the reverse transcription process can be delayed by using the reverse transcriptase inhibitor nevirapine. This study is of significant importance for the development of antiviral drugs.
Reverse transcription uses the reverse transcriptase enzyme to synthesize deoxyribonucleic acid (DNA) from a ribonucleic acid (RNA) template. This plays an essential role in viral replication. There are still, however, many unknown facts regarding the timing and dynamic processes involved in this life stage. Here, three types of dual-fluorescence human immunodeficiency virus type-1 (HIV-1) particles were constructed with high infectivity, and the sequential process of reverse transcription was observed by real-time imaging of a single HIV-1 particle. Viral uncoating occurred at 60-120 min post infection. Subsequently, at 120-180 min post infection, the viral genome was separated into two parts and reverse-transcribed to generate a DNA product. Nevirapine (NVP), a reverse transcriptase inhibitor, can delay the dynamic process. This study revealed a delicate, sequential, and complex relationship between uncoating and reverse transcription, which may facilitate the development of antiviral drugs.
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