Constraints in simian virus 40 (SV40) encapsidation, as determined by SV40-based shuttle viruses.
Simian virus 40 (SV40)-based shuttle vectors, containing the SV40 late genes, can be packaged as infectious pseudovirions. In terms of their function as bacterial plasmids, modifications in the overall size of these plasmids can be tolerated within a very wide range, which has allowed us to determine the requirements for SV40 encapsidation, free of the more stringent limitations of SV40 virus. Monkey COS7 cells were transfected with over- and undersized SV40-based shuttle virus plasmids and their progeny have been analysed to follow the stability and evolution of these genomes. Two of the three plasmids analysed undergo recombination, generating molecules with sizes of between 4.0 and to 4.8 kb which were selected after multiple lytic cycles. This size range may correspond to the DNA lengths preferentially packaged in SV40 capsids. The structure of the rearranged plasmids indicates that there is a strong selective pressure for genomes that retain the functions necessary for replication and virus production. Depending on the parent DNA, two main classes of rearrangements were generated: duplications in tandem with the SV40 origin of replication and deletions. Both classes are probably a result of selective size and replicative advantages, which are then biologically amplified during plasmid transmission as virus particles.
Menck
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CF
Laboratory of Molecular Mutagenesis, Institut de Recherches Scientifiques sur le Cancer, Villejuif, France.
James
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M
Benoit
A
A
Sarasin
A
A
D000818:Animals; D015139:Blotting, Southern; D002213:Capsid / Q000235:genetics*; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics; D015247:Deoxyribonuclease HindIII; D004587:Electrophoresis, Agar Gel; D015321:Gene Rearrangement; D005822:Genetic Vectors; D010957:Plasmids; D015183:Restriction Mapping; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection; D014771:Virion / Q000235:genetics*; D014779:Virus Replication
D004279:DNA, Viral; D015247:Deoxyribonuclease HindIII