Aronoff R, Linial M
Department of Microbiology, University of Washington, Seattle 98105.
J Virol. 1991 Jan;65(1):71-80. doi: 10.1128/JVI.65.1.71-80.1991.
Encapsidation of retroviral RNA has been shown to be dependent on specific cis-acting signals, in particular, the packaging region (psi) located near the 5' end of the retroviral genome. In this report, we show that a 683-base avian extended packaging sequence (psi+) derived from Rous sarcoma virus will direct packaging of heterologous hygromycin mRNA into avian virions when present at the 3' end of the transcript in the sense orientation. However, this packaging is not as efficient as the packaging of RNA encoded by a standard avian retroviral vector. A quail cell line containing a Rous sarcoma virus mutant, SE21Q1b, produces virions which will package endogenous cellular mRNAs randomly, roughly in proportion to their intracellular concentrations. We found that viral particles from SE21Q1b retain the capacity to specifically encapsidate hygromycin mRNAs containing the avian psi+. To determine whether packaging of cellular mRNA would occur in other retroviral packaging lines, we assayed virion RNA isolated from the retroviral particles produced by avian and murine packaging lines for the presence of endogenous cellular mRNAs. Endogenous cellular mRNAs were not found randomly packaged into virions produced by any of the packaging lines examined except SE21Q1b. Some specific sequences, however, were found packaged into avian virions. Endogenous retrovirus-related mink cell focus-inducing murine leukemia virus RNAs and 30S viruslike RNAs were found to be efficiently packaged into murine virions even in the presence of RNAs containing all cis-acting retroviral sequences.
逆转录病毒RNA的包装已被证明依赖于特定的顺式作用信号,特别是位于逆转录病毒基因组5'端附近的包装区域(ψ)。在本报告中,我们表明,源自劳氏肉瘤病毒的683个碱基的禽类延伸包装序列(ψ+),当以正义方向存在于转录本的3'端时,将指导异源潮霉素mRNA包装到禽病毒粒子中。然而,这种包装效率不如标准禽逆转录病毒载体编码的RNA的包装效率高。含有劳氏肉瘤病毒突变体SE21Q1b的鹌鹑细胞系产生的病毒粒子会随机包装内源性细胞mRNA,大致与其细胞内浓度成比例。我们发现,来自SE21Q1b的病毒粒子保留了特异性包装含有禽ψ+的潮霉素mRNA的能力。为了确定细胞mRNA的包装是否会在其他逆转录病毒包装细胞系中发生,我们检测了从禽和鼠包装细胞系产生的逆转录病毒粒子中分离的病毒粒子RNA中是否存在内源性细胞mRNA。除了SE21Q1b外,在所检测的任何包装细胞系产生的病毒粒子中均未发现内源性细胞mRNA被随机包装。然而,发现一些特定序列被包装到禽病毒粒子中。即使在存在含有所有顺式作用逆转录病毒序列的RNA的情况下,内源性逆转录病毒相关的貂细胞灶诱导型鼠白血病病毒RNA和30S病毒样RNA也被发现有效地包装到鼠病毒粒子中。