Biologycal Physics Laboratory, Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí 78000, Mexico.
Molecular Biology Division, IPICYT, Instituto Potosino de Investigación Científica y Tecnológica A.C., San Luis Potosí 78216, Mexico.
Viruses. 2023 Oct 4;15(10):2046. doi: 10.3390/v15102046.
(CCMV) and (BMV) are naked plant viruses with similar characteristics; both form a T = 3 icosahedral protein capsid and are members of the family. It is well known that these viruses completely disassemble and liberate their genome at a pH around 7.2 and 1 M ionic strength. However, the 1 M ionic strength condition is not present inside cells, so an important question is how these viruses deliver their genome inside cells for their viral replication. There are some studies reporting the swelling of the CCMV virus using different techniques. For example, it is reported that at a pH~7.2 and low ionic strength, the swelling observed is about 10% of the initial diameter of the virus. Furthermore, different regions within the cell are known to have different pH levels and ionic strengths. In this work, we performed several experiments at low ionic strengths of 0.1, 0.2, and 0.3 and systematically increased the pH in 0.2 increments from 4.6 to 7.4. To determine the change in virus size at the different pHs and ionic strengths, we first used dynamic light scattering (DLS). Most of the experiments agree with a 10% capsid swelling under the conditions reported in previous works, but surprisingly, we found that at some particular conditions, the virus capsid swelling could be as big as 20 to 35% of the original size. These measurements were corroborated by atomic force microscopy (AFM) and transmission electron microscopy (TEM) around the conditions where the big swelling was determined by DLS. Therefore, this big swelling could be an easier mechanism that viruses use inside the cell to deliver their genome to the cell machinery for viral replication.
(CCMV)和(BMV)是具有相似特征的裸露植物病毒;两者都形成 T = 3 二十面体蛋白衣壳,是 家族的成员。众所周知,这些病毒在 pH 值约为 7.2 和 1 M 离子强度下完全解体并释放其基因组。然而,细胞内不存在 1 M 离子强度条件,因此一个重要的问题是这些病毒如何将其基因组递送到细胞内以进行病毒复制。有一些研究报告使用不同的技术报告了 CCMV 病毒的肿胀。例如,据报道,在 pH~7.2 和低离子强度下,观察到的肿胀约为病毒初始直径的 10%。此外,已知细胞内的不同区域具有不同的 pH 值和离子强度。在这项工作中,我们在低离子强度为 0.1、0.2 和 0.3 的条件下进行了几项实验,并系统地将 pH 值从 4.6 以 0.2 的增量增加到 7.4。为了确定在不同 pH 值和离子强度下病毒大小的变化,我们首先使用动态光散射(DLS)。大多数实验与先前工作中报道的条件下 10%的衣壳肿胀一致,但令人惊讶的是,我们发现,在某些特定条件下,病毒衣壳的肿胀可能高达原始尺寸的 20%到 35%。这些测量结果通过原子力显微镜(AFM)和透射电子显微镜(TEM)得到了证实,这些显微镜是在通过 DLS 确定大肿胀的条件下进行的。因此,这种大肿胀可能是病毒在细胞内使用的一种更容易的机制,可将其基因组递送到细胞机制以进行病毒复制。