Xing Li, Tikoo Suresh K
Vaccine & Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Virol J. 2009 Feb 5;6:16. doi: 10.1186/1743-422X-6-16.
Earlier, we detected viral RNAs packaged in the porcine adenovirus (PAdV) -3 virions. Using Southern blot analysis, we further demonstrated that the viral RNAs were predominantly packaged in CsCl purified mature capsids (containing viral genome) than empty/intermediate capsids. Some of the packaged viral RNAs appear to be polyadenylated. Real-time reverse transcription (RT)-PCR analysis indicated that the copy number of the tested viral mRNAs encoding E1Bsmall and fiber proteins was less than one per full capsid. Moreover, detection of viral RNA packaged in CsCl purified human adenovirus (HAdV) -5 virions indicates that the viral RNA packaging might be a common phenomenon in members of Adenoviridae family. Further quantitative analysis of viral protein, DNA, and RNA in CsCl purified mature and empty/intermediate capsids of recombinant HAdV-5 expressing enhanced green fluorescent protein indicated that the traceable viral RNA detected in empty/intermediate capsids seems associated with the presence of traceable viral genomic DNA. Taken together, our data suggest that the viral RNAs may be passively packaged in adenovirus virion during encapsidation of viral genomic DNA in cell nuclei. Thus, viral RNA packaging may be a characteristic feature of adenoviral genomic DNA encapsidation.
此前，我们检测到包装在猪腺病毒（PAdV）-3病毒粒子中的病毒RNA。通过Southern印迹分析，我们进一步证明，与空衣壳/中间衣壳相比，病毒RNA主要包装在CsCl纯化的成熟衣壳（含有病毒基因组）中。一些包装的病毒RNA似乎是聚腺苷酸化的。实时逆转录（RT）-PCR分析表明，编码E1B小蛋白和纤维蛋白的测试病毒mRNA的拷贝数在每个完整衣壳中少于一个。此外，对CsCl纯化的人腺病毒（HAdV）-5病毒粒子中包装的病毒RNA的检测表明，病毒RNA包装可能是腺病毒科成员中的一种常见现象。对表达增强型绿色荧光蛋白的重组HAdV-5的CsCl纯化的成熟衣壳和空衣壳/中间衣壳中的病毒蛋白、DNA和RNA进行进一步定量分析表明，在空衣壳/中间衣壳中检测到的可追踪病毒RNA似乎与可追踪病毒基因组DNA的存在有关。综上所述，我们的数据表明，在细胞核中病毒基因组DNA衣壳化过程中，病毒RNA可能被被动包装在腺病毒病毒粒子中。因此，病毒RNA包装可能是腺病毒基因组DNA衣壳化的一个特征。
