Release of Genotype 1 Hepatitis E Virus from Cultured Hepatoma and Polarized Intestinal Cells Depends on Open Reading Frame 3 Protein and Requires an Intact PXXP Motif
Release of Genotype 1 Hepatitis E Virus from Cultured Hepatoma and Polarized Intestinal Cells Depends on Open Reading Frame 3 Protein and Requires an Intact PXXP Motif
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DOI：
10.1128/jvi.00593-10
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发表时间：
2010-09-15
影响因子：
5.4
通讯作者：
Purcell, Robert H.
中科院分区：
文献类型：
--
作者：
Emerson, Suzanne U.;Nguyen, Hanh T.;Purcell, Robert H.
戊型肝炎病毒基因型1毒株Sar55在已转染体外转录病毒基因组的亚克隆Caco-2肠细胞和Huh7肝癌细胞中复制，戊型肝炎病毒颗粒被释放到两种细胞系的培养基中。病毒从细胞中的排出取决于开放阅读框 3 (ORF3) 蛋白，而 ORF3 中富含脯氨酸的序列对于从培养细胞中排出和感染猕猴非常重要。细胞内 ORF3 蛋白积累和病毒释放均发生在极化 Caco-2 细胞的顶膜上。 ORF3蛋白和脂质与任一细胞系中产生的病毒颗粒密切相关； ORF2 表位被这些颗粒掩盖，不能用抗 ORF2 进行免疫沉淀。
Hepatitis E virus genotype 1 strain Sar55 replicated in subcloned Caco-2 intestinal cells and Huh7 hepatoma cells that had been transfected with in vitro transcribed viral genomes, and hepatitis E virions were released into the culture medium of both cell lines. Virus egress from cells depended on open reading frame 3 (ORF3) protein, and a proline-rich sequence in ORF3 was important for egress from cultured cells and for infection of macaques. Both intracellular ORF3 protein accumulation and virus release occurred at the apical membrane of polarized Caco-2 cells. ORF3 protein and lipids were intimately associated with virus particles produced in either cell line; ORF2 epitopes were masked in these particles and could not be immunoprecipitated with anti-ORF2.