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Analysis of complex mutations induced in cells by herpes simplex virus type-1
1Department of Microbiology, University of Texas Health Science Center, Houston 77225.
Virology
|April 1, 1991
Summary
Herpes simplex virus type-1 (HSV-1) infection stimulates nonhomologous recombination in cells, leading to complex plasmid mutations. Inserted DNA sequences were cellular, not viral, suggesting HSV-1 may mediate rearrangements of host DNA.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- Shuttle vector plasmids are crucial tools for studying DNA mutagenesis and recombination in mammalian cells.
- Herpes simplex virus type-1 (HSV-1) is a human DNA virus known to interact with host cell DNA processes.
- Understanding viral-mediated DNA alterations is key to comprehending viral pathogenesis and host-pathogen interactions.
Purpose of the Study:
- To investigate complex mutations in shuttle vector plasmids within COS-1 cells, comparing uninfected cells with those infected by HSV-1.
- To characterize the nature and origin of inserted DNA sequences found in enlarged plasmids from HSV-1 infected cells.
- To determine if HSV-1 infection influences recombination events and the types of mutations occurring in plasmid DNA.
Main Methods:
- Shuttle vector plasmid pZ189 was used as a target for mutagenesis in COS-1 cells.
- Plasmids were recovered from both noninfected and HSV-1 infected cells for analysis.
- Restriction enzyme digestion, DNA sequencing, and hybridization techniques were employed to map and identify mutations and inserted sequences.
Main Results:
- Complex mutations, including duplications and rearrangements, were observed in noninfected cells.
- In HSV-1 infected cells, 8 out of 15 plasmids showed enlarged sizes due to insertions of cellular DNA, ranging from 118 bp to 4500 bp.
- Inserted DNA sequences were confirmed as non-viral, hybridized to cellular DNA, and contained repeat sequences (e.g., alpha satellite, Alu, Kpn); junctions lacked transposable element features.
Conclusions:
- Herpes simplex virus type-1 (HSV-1) infection appears to stimulate nonhomologous recombination between DNA sequences in infected cells.
- The study suggests that cellular DNA sequences may serve as targets for virus-mediated genomic rearrangements.
- HSV-1 infection can lead to significant alterations in plasmid DNA structure through the insertion of host DNA.