Kinetics of porcine circovirus type 2 replication.
The kinetics of porcine circovirus type 2 (PCV2) replication in PK15 cells was examined. During productive infection, viral antigens, RNA transcripts and progeny viruses all increased in a time dependent manner. Viral antigens were observed in a few cells at 18 hour postinfection (h p.i.) and cell-free progeny viruses began to appear at about 30 h p.i. Viral transcripts were detected by 18 h p.i. and the capsid protein RNA of 950 nucleotides (nt) was the most abundant RNA species. Two other RNAs of sizes 750 and 450 nt, derived from the predicted replication associated protein (Rep) gene region, were also detected. These two RNAs share 3' common nucleotide sequences and they are transcribed in the same orientation as the proposed unspliced Rep RNA or the recently described Rep' RNA. The 35 kD capsid protein was observed at 30 h p.i. by Western blot analysis and it appeared to be the most immunodominant protein in swine exposed to PCV2. The capsid proteins of PCV type 1 and PCV2 each contain a nuclear localization signal sequence capable of targeting a reporter protein to the nucleoli of transfected cells when the capsid proteins were expressed as 3' fusion polypeptides. Although previous reports indicated that PCV2 capsid proteins localized predominantly in the nuclei of infected cells, we observed an abundant amount of PCV2 capsid proteins in the cytoplasm of many cells of the infected cultures. The cells that exhibited cytoplasmic capsid proteins also contained virus nucleic acids, indicating that these proteins were synthesized by the infected cells and not through uptake from the culture medium. Elucidation of the changes that affect the localization pattern of PCV2 capsid proteins, nuclear versus cytoplasmic, requires further investigation.
Cheung
A K
AK
Virus and Prion Diseases of Livestock Research Unit, National Animal Disease Center, USDA, Agricultural Research Service, Ames, Iowa 50010, USA. acheung@nadc.ars.usda.gov
Bolin
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SR
D000818:Animals; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D017925:Circovirus / Q000235:genetics / Q000502:physiology*; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology; D017403:In Situ Hybridization; D007700:Kinetics; D012367:RNA, Viral / Q000096:biosynthesis; D013552:Swine; D014162:Transfection; D014779:Virus Replication
D012367:RNA, Viral