HIV-1 Nucleocapsid Protein Binds Double-Stranded DNA in Multiple Modes to Regulate Compaction and Capsid Uncoating.
HIV-1 Nucleocapsid Protein Binds Double-Stranded DNA in Multiple Modes to Regulate Compaction and Capsid Uncoating.
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DOI：
10.3390/v14020235
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发表时间：
2022-01-25
期刊：
影响因子：
--
通讯作者：
Williams MC
中科院分区：
文献类型：
--
作者：
Gien H;Morse M;McCauley MJ;Kitzrow JP;Musier-Forsyth K;Gorelick RJ;Rouzina I;Williams MC
HIV-1核衣壳蛋白（NC）是病毒复制所必需的多功能蛋白。最近的研究表明，逆转录发生在完全完整的病毒衣壳内，逆转录的时间和脱壳是相关的。近10 kbp的病毒DNA基因组如何稳定地包含在直径与双链（ds）DNA的持久长度相似的狭窄衣壳内，以及NC在此过程中的作用还没有很好地理解。在这项研究中，我们使用光镊，荧光成像和原子力显微镜观察NC结合一个单一的长DNA基板在多个模式。我们发现，NC结合和饱和的DNA底物的非特异性结合模式，触发均匀的DNA自吸引力，凝聚成一个紧密的小球在一个恒定的力高达10 pN的DNA。当NC从溶液中去除时，小球随时间消散，但特异性结合的NC保持较不紧凑但高度稳定的长距离DNA环。另外，使用AFM成像观察到两种结合模式。这些结果表明NC紧凑DNA的多种结合模式与逆转录相容的构象，调节衣壳上的基因组压力并防止过早脱壳。
The HIV-1 nucleocapsid protein (NC) is a multi-functional protein necessary for viral replication. Recent studies have demonstrated reverse transcription occurs inside the fully intact viral capsid and that the timing of reverse transcription and uncoating are correlated. How a nearly 10 kbp viral DNA genome is stably contained within a narrow capsid with diameter similar to the persistence length of double-stranded (ds) DNA, and the role of NC in this process, are not well understood. In this study, we use optical tweezers, fluorescence imaging, and atomic force microscopy to observe NC binding a single long DNA substrate in multiple modes. We find that NC binds and saturates the DNA substrate in a non-specific binding mode that triggers uniform DNA self-attraction, condensing the DNA into a tight globule at a constant force up to 10 pN. When NC is removed from solution, the globule dissipates over time, but specifically-bound NC maintains long-range DNA looping that is less compact but highly stable. Both binding modes are additionally observed using AFM imaging. These results suggest multiple binding modes of NC compact DNA into a conformation compatible with reverse transcription, regulating the genomic pressure on the capsid and preventing premature uncoating.
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影响因子：
11.7
作者：
Chaurasiya, Kathy R.;Paramanathan, Thayaparan;McCauley, Micah J.;Williams, Mark C.
通讯作者：
Williams, Mark C.
影响因子：
64.5
作者：
Kessl, Jacques J.;Kutluay, Sebla B.;Townsend, Dana;Rebensburg, Stephanie;Slaughter, Alison;Larue, Ross C.;Shkriabai, Nikoloz;Bakouche, Nordine;Fuchs, James R.;Bieniasz, Paul D.;Kvaratskhelia, Mamuka
通讯作者：
Kvaratskhelia, Mamuka
DOI：
10.3390/v12111234
发表时间：
2020-10-30
期刊：
Viruses
影响因子：
--
作者：
Francis AC;Marin M;Prellberg MJ;Palermino-Rowland K;Melikyan GB
通讯作者：
Melikyan GB
影响因子：
8.6
作者：
Kornyshev, AA;Leikin, S
通讯作者：
Leikin, S
影响因子：
6.7
作者：
Brandariz-Nunez, Alberto;Robinson, Scott J.;Evilevitch, Alex
通讯作者：
Evilevitch, Alex