Modification of the trypsin cleavage site of rotavirus VP4 to a furin-sensitive form does not enhance replication efficiency.
Modification of the trypsin cleavage site of rotavirus VP4 to a furin-sensitive form does not enhance replication efficiency.
复制标题
将轮状病毒 VP4 的胰蛋白酶切割位点修饰为弗林蛋白酶敏感形式并不会提高复制效率。
DOI：
10.1099/vir.0.033886-0
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
K. Taniguchi
中科院分区：
文献类型：
--
作者：
S. Komoto;M. Wakuda;Tomihiko Ide;Gen Niimi;Y. Maeno;Kyoko Higo;K. Taniguchi
轮状病毒 (RV) 的感染性取决于其刺突蛋白 VP4 的蛋白水解裂解所触发的激活过程。这种激活裂解是由体内肠腔中的外源胰蛋白酶进行的。在这里，我们报告了表达 cDNA 衍生的 VP4 的重组 RV 的生成和表征，该 RV 具有可被内源性弗林蛋白酶和外源性胰蛋白酶识别的修饰切割位点（位置 247 处的精氨酸）。出乎意料的是，突变病毒（KU//rVP4-R247Furin）在没有外源蛋白酶的情况下无法形成噬菌斑，尽管病毒粒子上的突变VP4被内源弗林蛋白酶有效裂解。此外，与携带真实 VP4 (KU//rVP4) 的亲本病毒相比，即使在胰蛋白酶存在下，KU//rVP4-R247Furin 在 MA104 和 CV-1 细胞中的感染性也受损。尽管KU//rVP4-R247Furin的总滴度与KU//rVP4相当，但与KU//rVP4相比，KU//rVP4-R247Furin的细胞外滴度明显低于其细胞相关滴度。相比之下，这两种病毒在弗林蛋白酶缺陷型 LoVo 细胞系中表现出相似的生长。这些结果表明，弗林蛋白酶对 VP4 的细胞内裂解可能不利于 RV 感染，可能是由于病毒释放过程效率低下。
The infectivity of rotavirus (RV) is dependent on an activation process triggered by the proteolytic cleavage of its spike protein VP4. This activation cleavage is performed by exogenous trypsin in the lumen of the intestines in vivo. Here, we report the generation and characterization of a recombinant RV expressing cDNA-derived VP4 with a modified cleavage site (arginine at position 247) recognized by endogenous furin as well as exogenous trypsin. Unexpectedly, the mutant virus (KU//rVP4-R247Furin) was incapable of plaque formation without an exogenous protease, although the mutant VP4s on virions were efficiently cleaved by endogenous furin. Furthermore, KU//rVP4-R247Furin showed impaired infectivity in MA104 and CV-1 cells even in the presence of trypsin compared with the parental virus carrying authentic VP4 (KU//rVP4). Although the total titre of KU//rVP4-R247Furin was comparable to that of KU//rVP4, the extracellular titre of KU//rVP4-R247Furin was markedly lower than its cell-associated titre in comparison with that of KU//rVP4. In contrast, the two viruses showed similar growth in a furin-defective LoVo cell line. These results suggest that intracellular cleavage of VP4 by furin may be disadvantageous for RV infectivity, possibly due to an inefficient virus release process.
轮状病毒VP4的体内裂解。
DOI：
10.1099/0022-1317-77-3-391
发表时间：
1996
期刊：
The Journal of general virology.
影响因子：
--
作者：
Ludert,JE;Krishnaney,AA;Burns,JW;Vo,PT;Greenberg,HB
通讯作者：
Greenberg,HB