Back to Search
Start Over
Human Hepatitis B Virus Production in Avian Cells Is Characterized by Enhanced RNA Splicing and the Presence of Capsids Containing Shortened Genomes
- Source :
- PLoS ONE, PLoS ONE, Vol 7, Iss 5, p e37248 (2012)
- Publication Year :
- 2012
- Publisher :
- Public Library of Science, 2012.
-
Abstract
- Experimental studies on hepatitis B virus (HBV) replication are commonly done with human hepatoma cells to reflect the natural species and tissue tropism of the virus. However, HBV can also replicate, upon transfection of virus coding plasmids, in cells of other species. In such cross-species transfection experiments with chicken LMH hepatoma cells, we previously observed the formation of HBV genomes with aberrant electrophoretic mobility, in addition to the those DNA species commonly seen in human HepG2 hepatoma cells. Here, we report that these aberrant DNA forms are mainly due to excessive splicing of HBV pregenomic RNA and the abundant synthesis of spliced DNA products, equivalent to those also made in human cells, yet at much lower level. Mutation of the common splice acceptor site abolished splicing and in turn enhanced production of DNA from full-length pgRNA in transfected LMH cells. The absence of splicing made other DNA molecules visible, that were shortened due to the lack of sequences in the core protein coding region. Furthermore, there was nearly full-length DNA in the cytoplasm of LMH cells that was not protected in viral capsids. Remarkably, we have previously observed similar shortened genomes and non-protected viral DNA in human HepG2 cells, yet exclusively in the nucleus where uncoating and final release of viral genomes occurs. Hence, two effects reflecting capsid disassembly in the nucleus in human HepG2 cells are seen in the cytoplasm of chicken LMH cells.
- Subjects :
- Hepatitis B virus
viruses
RNA Splicing
lcsh:Medicine
Viral diseases
Virus Replication
Biochemistry
Microbiology
Polymerase Chain Reaction
Viral Packaging
Hepatitis
Capsid
Virology
Cell Line, Tumor
Molecular Cell Biology
Viral Nucleic Acid
Animals
Humans
lcsh:Science
Biology
DNA Primers
lcsh:R
DNA
Hepatitis B
Viral Replication
Nucleic acids
Blotting, Southern
RNA processing
RNA
Medicine
Infectious diseases
lcsh:Q
Chickens
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 7
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.pmid.dedup....d366b5fdaa2a7e2d1188bfe6eea4683b