Directly monitoring individual retrovirus budding events using atomic force microscopy
Directly monitoring individual retrovirus budding events using atomic force microscopy
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DOI：
10.1529/biophysj.107.114579
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发表时间：
2008-01-01
影响因子：
3.4
通讯作者：
Rousso, Itay
中科院分区：
文献类型：
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作者：
Gladnikoff, Micha;Rousso, Itay
逆转录病毒出芽是病毒复制周期中的关键步骤。尽管如此，人们对出芽的基本机制知之甚少，主要是由于技术限制，无法在真实的时间内观察出芽的形成。能够监测出芽动态的方法遭受分辨率不足，而其他方法，如电子显微镜，不具有在生理条件下操作的能力。在这里，我们应用原子力显微镜实时可视化个别莫洛尼小鼠白血病病毒萌芽事件。通过使用单粒子分析方法，我们能够观察到不同的模式，在萌芽，否则保持透明。我们发现，芽的形成遵循至少两个动力学不同的途径。大多数病毒粒子（74%）是在缓慢过程（>45分钟）中产生的，其余粒子（26%）通过快速过程（
Retrovirus budding is a key step in the virus replication cycle. Nonetheless, very little is known about the underlying mechanism of budding, primarily due to technical limitations preventing visualization of bud formation in real time. Methods capable of monitoring budding dynamics suffer from insufficient resolution, whereas other methods, such as electron microscopy, do not have the ability to operate under physiological conditions. Here we applied atomic force microscopy to real-time visualization of individual Moloney murine leukemia virus budding events. By using a single-particle analysis approach, we were able to observe distinct patterns in budding that otherwise remain transparent. We find that bud formation follows at least two kinetically distinct pathways. The majority of virions (74%) are produced in a slow process (>45 min), and the remaining particles (26%) assemble via a fast process (