Herpesvirus-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication.

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Abstract
The genome of herpes simplex virus 1 or 2 consists of 2 components, L and S, which invert relative to each other during infection. As a result, viral DNA consists of 4 equimolar populations of molecules differing solely in the relative orientations of the L and S components. The a sequences, located in the same orientation at the genomic termini and in inverted orientation at the L-S junction, were previously shown play a key role in the inversion of L and S components. A virus-dependent system designed to allow identification of the viral genes capable of acting in trans to invert DNA flanked by inverted copies of a sequences is described. In this system, African green monkey kidney Vero cells are converted to the thymidine kinase-positive phenotype with a chimeric plasmid carrying the thymidine kinase gene flanked by inverted copies of the a sequence and linked to an origin of viral DNA replication derived from the S component. The DNA introduced into the cells is retained and propagated in its original sequence arrangement as head-to-tail concatemers. Infection of these cells with herpes simplex virus 1 or 2 results in as much as 100-fold amplification of the plasmid sequences and inversion of the DNA flanked by copies of the a sequence. In infected cells, the amplified resident DNA accumulates in head-to-tail concatemers and no rearrangement other than the inversions could be detected. Thus, the a sequence-dependent inversions apparently require trans-acting viral gene products.

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