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Papers In Press, published online ahead of print November 7, 2000
J. Biol. Chem, 10.1074/jbc.M005627200
Submitted on June 27, 2000
Revised on November 6, 2000
Accepted on November 6, 2000
Medical Biochemistry, Goteborg University, Goteborg SE 405 30
Corresponding Author: per.elias{at}medkem.gu.se
Homologous recombination was examined in cells infected with Herpes simplex virus type I. Circular and linear DNA with directly repeated sequences was introduced as recombination substrates into cells. Recombination was measured either by origin-dependent amplification of recombination products or by recombination-dependent expression of luciferase from a disrupted gene. Homologous recombination in BHK cells converted linear DNA to circular templates for DNA replication and luciferase expression in the complete absence of virus. The products of homologous recombination were efficiently amplified by the viral replication apparatus. The efficiency of recombination was dependent on the structure of the substrate as well as the cell type. Linear DNA with the direct repeats at internal positions failed to recombine in Balb/c 3T3 cells and induced p53 dependent apoptosis . In contrast, linear DNA with directly repeated sequences precisely at the ends recombined and replicated in 3T3 cells. Homologous recombination in BHK cells did not depend on the position of the repeated sequences. We conclude that homologous recombination is independent of viral gene functions and it is likely to be carried out by cellular proteins. We suggest that homologous recombination between directly repeated sequences in the linear HSV-1 virus chromosome may help to avoid p53 dependent apoptosis and to promote viral DNA replication.
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