Abstract
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.
Highlights
The phenomenon of encapsidation of viral RNAs was initially uncovered in members of Herpesviridae family [1,2,3,4]
We examined the incorporation of viral RNAs in mature and empty/intermediate capsids of porcine adenovirus (PAdV)-3 and human adenovirus (HAdV)-5
Preparation and characterization of PAdV-3 capsids At late times during adenovirus infection, two kinds of particles are abundant, which can be separated by CsCl gradient centrifugation [22]
Summary
The phenomenon of encapsidation of viral RNAs was initially uncovered in members of Herpesviridae family [1,2,3,4] These RNAs could be translated into proteins that would function prior to de novo transcription from the viral genome [1,2,5]. Studies with herpes simplex virus (HSV) -1 and Kaposi's sarcoma-associated herpesvirus (KSHV) suggested that some virion RNAs were incorporated into the virions [1,4]. Adenoviruses are another family of DNA viruses that infect a wide variety of mammals and birds [9]. The characteristics of packaged viral RNAs were further examined by Southern blot hybridization and real-time RT-PCR analysis
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