Serghini M A, Fuchs M, Pinck M, Reinbolt J, Walter B, Pinck L
Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg, France.
J Gen Virol. 1990 Jul;71 ( Pt 7):1433-41. doi: 10.1099/0022-1317-71-7-1433.
The nucleotide sequence of the genomic RNA2 (3774 nucleotides) of grapevine fanleaf virus strain F13 was determined from overlapping cDNA clones and its genetic organization was deduced. Two rapid and efficient methods were used for cDNA cloning of the 5' region of RNA2. The complete sequence contained only one long open reading frame of 3555 nucleotides (1184 codons, 131K product). The analysis of the N-terminal sequence of purified coat protein (CP) and identification of its C-terminal residue have allowed the CP cistron to be precisely positioned within the polyprotein. The CP produced by proteolytic cleavage at the Arg/Gly site between residues 680 and 681 contains 504 amino acids (Mr 56019) and has hydrophobic properties. The Arg/Gly cleavage site deduced by N-terminal amino acid sequence analysis is the first for a nepovirus coat protein and for plant viruses expressing their genomic RNAs by polyprotein synthesis. Comparison of GFLV RNA2 with M RNA of cowpea mosaic comovirus and with RNA2 of two closely related nepoviruses, tomato black ring virus and Hungarian grapevine chrome mosaic virus, showed strong similarities among the 3' non-coding regions but less similarity among the 5' end non-coding sequences than reported among other nepovirus RNAs.
通过重叠cDNA克隆确定了葡萄扇叶病毒F13株系基因组RNA2（3774个核苷酸）的核苷酸序列，并推导了其基因组织结构。使用了两种快速有效的方法对RNA2的5'区域进行cDNA克隆。完整序列仅包含一个3555个核苷酸的长开放阅读框（1184个密码子，产物为131K）。对纯化衣壳蛋白（CP）的N端序列分析及其C端残基的鉴定，使得CP顺反子能够精确地定位在多聚蛋白中。在第680和681位残基之间的Arg/Gly位点通过蛋白水解切割产生的CP含有504个氨基酸（Mr 56019），具有疏水特性。通过N端氨基酸序列分析推导的Arg/Gly切割位点是线虫传多面体病毒衣壳蛋白以及通过多聚蛋白合成表达其基因组RNA的植物病毒中的首个此类位点。将葡萄扇叶病毒RNA2与豇豆花叶病毒的M RNA以及两种密切相关的线虫传多面体病毒番茄黑环病毒和匈牙利葡萄铬花叶病毒的RNA2进行比较，结果显示3'非编码区之间有很强的相似性，但5'端非编码序列之间相似度低于其他线虫传多面体病毒RNA之间报道的相似度。
