Marmey P, Bothner B, Jacquot E, de Kochko A, Ong C A, Yot P, Siuzdak G, Beachy R N, Fauquet C M
Plant Division, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California, 92037, USA.
Virology. 1999 Jan 20;253(2):319-26. doi: 10.1006/viro.1998.9519.
Rice tungro bacilliform virus (RTBV) is a plant pararetrovirus and a member of the Caulimoviridae family and closely related to viruses in the Badnavirus genus. The coat protein of RTBV is part of the large polyprotein encoded by open reading frame 3 (ORF3). ORF3 of an RTBV isolate from Malaysia was sequenced (accession no. AF076470) and compared with published sequences for the region that encodes the coat protein or proteins. Molecular mass of virion proteins was determined by mass spectrometry (matrix-assisted laser desorption/ionization-TOF) performed on purified virus particles from three RTBV isolates from Malaysia. The N- and C-terminal amino acid sequences of the coat protein were deduced from the mass spectral analysis, leading to the conclusion that purified virions contain a single coat protein of 37 kDa. The location of the coat protein domain in ORF3 was reinforced as a result of immunodetection reactions using antibodies raised against six different segments of ORF3 using Western immunoblots after SDS-PAGE and isoelectrofocusing of proteins purified from RTBV particles. These studies demonstrate that RTBV coat protein is released from the polyprotein as a single coat protein of 37 kDa.
水稻东格鲁杆状病毒(RTBV)是一种植物类逆转录病毒,属于花椰菜花叶病毒科,与杆状DNA病毒属的病毒密切相关。RTBV的外壳蛋白是由开放阅读框3(ORF3)编码的大的多聚蛋白的一部分。对来自马来西亚的一株RTBV分离株的ORF3进行了测序(登录号AF076470),并与已发表的编码外壳蛋白区域的序列进行了比较。通过对来自马来西亚的三株RTBV分离株的纯化病毒颗粒进行质谱分析(基质辅助激光解吸/电离飞行时间质谱)来确定病毒粒子蛋白的分子量。通过质谱分析推导了外壳蛋白的N端和C端氨基酸序列,得出纯化的病毒粒子含有一种37 kDa的单一外壳蛋白的结论。在对从RTBV颗粒中纯化的蛋白质进行SDS-PAGE和等电聚焦后,利用针对ORF3六个不同片段产生的抗体进行免疫检测反应(Western免疫印迹法),进一步确定了外壳蛋白结构域在ORF3中的位置。这些研究表明,RTBV外壳蛋白作为一种37 kDa的单一外壳蛋白从多聚蛋白中释放出来。