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利用不同物理化学线索探索细小病毒单个颗粒中的核酸凝聚和释放情况。

Exploring nucleic acid condensation and release from individual parvovirus particles with different physicochemical cues.

作者信息

Strobl K, Mateu M G, de Pablo Pedro J

机构信息

Department of Condensed Matter Physics Universidad Autónoma de Madrid, 28049, Madrid, Spain.

Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, 28049, Madrid, Spain.

出版信息

Virology. 2023 Apr;581:1-7. doi: 10.1016/j.virol.2023.01.010. Epub 2023 Feb 14.

Abstract

In the infection cycle, viruses release their genome in the host cell during uncoating. Here we use a variety of physicochemical procedures to induce and monitor the in vitro uncoating of ssDNA from individual Minute Virus of Mice (MVM) particles. Our experiments revealed two pathways of genome release: i) filamentous ssDNA appearing around intact virus particles when using gradual mechanical fatigue and heating at moderate temperature (50 °C). ii) thick structures of condensed ssDNA appearing when the virus particle is disrupted by mechanical nanoindentations, denaturing agent guanidinium chloride and high temperature (70 °C). We propose that in the case of filamentous ssDNA, when the capsid integrity is conserved, the genome is externalized through one channel of the capsid pores. However, the disruption of virus particles revealed a native structure of condensed genome. The mechanical analysis of intact particles after DNA strands ejection confirm the stabilization role of ssDNA in MVM.

摘要

在感染周期中,病毒在脱壳过程中将其基因组释放到宿主细胞中。在这里,我们使用各种物理化学方法来诱导和监测来自单个小鼠微小病毒(MVM)颗粒的单链DNA(ssDNA)的体外脱壳。我们的实验揭示了基因组释放的两条途径:i)当使用逐渐的机械疲劳和在中等温度(50°C)下加热时,丝状ssDNA出现在完整病毒颗粒周围。ii)当病毒颗粒被机械纳米压痕、变性剂氯化胍和高温(70°C)破坏时,出现浓缩ssDNA的厚结构。我们提出,在丝状ssDNA的情况下,当衣壳完整性得以保留时,基因组通过衣壳孔的一个通道外化。然而,病毒颗粒的破坏揭示了浓缩基因组的天然结构。DNA链射出后对完整颗粒的力学分析证实了ssDNA在MVM中的稳定作用。

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