首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Giardiavirus double-stranded RNA genome encodes a capsid polypeptide and a gag-pol-like fusion protein by a translation frameshift.
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Giardiavirus double-stranded RNA genome encodes a capsid polypeptide and a gag-pol-like fusion protein by a translation frameshift.

机译:贾第鞭毛虫病毒双链RNA基因组通过翻译移码编码衣壳多肽和gag-pol样融合蛋白。

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摘要

Giardiavirus is a small, nonenveloped virus comprising a monopartite double-stranded RNA genome, a major protein of 100 kDa, and a less abundant polypeptide of 190 kDa. It can be isolated from the culture supernatant of Giardia lamblia, a parasitic flagellate in human and other mammals, and efficiently infects other virus-free G. lamblia. A single-stranded copy of the viral RNA can be electroporated into uninfected G. lamblia cells to complete the viral replication cycle. Giardiavirus genomic cDNA of 6100 nt was constructed and its sequence revealed the presence of two large open reading frames that are separated by a -1 frameshift and share an overlap of 220 nt. The 3' open reading frame contains all consensus RNA-dependent RNA polymerase sequence motifs. A heptamer-pseudoknot structure similar to those found at ribosomal slippage sites in retroviruses and yeast killer virus was identified within this overlap. Immunostudies using antisera against synthesized peptides from four regions in the two open reading frames indicated that the 100- and 190-kDa viral proteins share a common domain in the amino-terminal region. But the 190-kDa protein makes a -1 switch of its reading frame beyond the presumed slippage heptamer and is therefore a -1 frameshift fusion protein similar to the gag-pol fusion protein found in retroviruses.
机译:贾第鞭毛虫病毒是一种小型无包膜病毒,包含单部分双链RNA基因组,100 kDa的主要蛋白质和190 kDa的较不丰富的多肽。它可以从贾第鞭毛虫的培养上清液中分离出来,贾第鞭毛虫是人和其他哺乳动物的寄生鞭毛,可以有效感染其他无病毒的兰氏鞭毛虫。可以将病毒RNA的单链副本电穿孔到未感染的G.lamblia细胞中,以完成病毒复制周期。构建了6100 nt的贾第鞭毛虫病毒基因组cDNA,其序列揭示了两个大的开放阅读框的存在,它们被-1移码分开并共有220 nt的重叠。 3'开放阅读框包含所有共有的RNA依赖性RNA聚合酶序列基序。在此重叠区域内鉴定出类似于在逆转录病毒和酵母杀伤病毒的核糖体滑脱位点发现的七聚体假单胞菌结构。使用抗血清针对来自两个开放阅读框中四个区域的合成肽进行的免疫研究表明,100kDa和190kDa的病毒蛋白在氨基末端区域共享一个公共域。但是190 kDa的蛋白质使其阅读框发生了-1转换,超出了预期的滑动七聚体,因此是-1移码融合蛋白,类似于逆转录病毒中的gag-pol融合蛋白。

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