Cleavage of rotavirus VP4 in vivo.
Cleavage of rotavirus VP4 in vivo.
复制标题
轮状病毒VP4的体内裂解。
DOI：
10.1099/0022-1317-77-3-391
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Greenberg,HB
中科院分区：
文献类型：
--
作者：
Ludert,JE;Krishnaney,AA;Burns,JW;Vo,PT;Greenberg,HB
轮状病毒颗粒的感染性依赖于外部衣壳蛋白VP 4在特定位点的蛋白水解裂解。该切割事件产生两个片段，鉴定为VP 5 * 和VP 8 *。假设当VP 4处于未切割状态时，颗粒更稳定，但无感染性。未裂解的VP 4和由此产生的增加的稳定性可能有利于病毒抵抗环境降解，直到它感染易感宿主。当VP 4在宿主胃肠道的内腔中被切割时，病毒颗粒将变得不稳定但更具感染性。为了验证这一假设，进行了一系列实验以分析由感染的宿主释放到环境中的病毒上的VP 4的切割状态。免疫印迹法的肠冲洗液来自感染的EDIM病毒（EW）或野生型鼠轮状病毒EDIM-Cambridge的毒力细胞培养适应的变种的婴儿和成年BALB/c小鼠进行了分析。这些样品中几乎所有的VP 4都是裂解形式。此外，对感染EW的幼崽经胰蛋白酶处理和未处理的肠道内容物进行细胞培养滴定表明，在添加胰蛋白酶之前，排泄的病毒已完全活化。还观察到，胰蛋白酶活化的病毒在初始动物中引发感染方面与含有完整VP 4的病毒体相比没有缺点。这些研究表明，VP 4在从肠细胞释放时被切割，并且脱落到环境中的病毒不具有完整的VP 4。
The infectivity of rotavirus particles is dependent on proteolytic cleavage of the outer capsid protein, VP4, at a specific site. This cleavage event yields two fragments, identified as VP5* and VP8*. It has been hypothesized that the particle is more stable, but non-infectious, when VP4 is in the uncleaved state. Uncleaved VP4 and the resultant increased stability might be advantageous for the virus to resist environmental degradation until it infects a susceptible host. When VP4 is cleaved in the lumen of the host’s gastrointestinal tract, the virus particle would become less stable but more infectious. To test this hypothesis, a series of experiments was undertaken to analyse the cleavage state of VP4 on virus shed by an infected host into the environment. Immunoblots of intestinal wash solutions derived from infant and adult BALB/c mice infected with a virulent cell culture-adapted variant of the EDIM virus (EW) or wild-type murine rotavirus EDIM-Cambridge were analysed. Virtually all of the VP4 in these samples was in the cleaved form. Moreover, cell culture titration of trypsintreated and untreated intestinal contents from pups infected with EW indicated that excreted virus is fully activated prior to trypsin addition. It was also observed that trypsin-activated virus has no disadvantage in initiating infection in naive animals over virions containing an intact VP4. These studies indicate that VP4 is cleaved upon release from the intestinal cell and that virus shed into the environment does not have an intact VP4.
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