Dating with herpes in columbus oh

These results suggest that increased mutation frequency alone cannot account for replication failure of most of the Exo domain mutations.We wished to examine the role of the 3′ to 5′ exo activity on the ability of the HSV-1 pol to modulate strand displacement synthesis in order to identify additional functions for this activity.We discuss the possibility that the lack of modulation of the strand displacement activity by the exo-deficient pol mutant may have catastrophic consequences for replication apart from a high spontaneous mutation frequency.Each of the HSV-1 genes encoding proteins used in these studies was cloned into a baculovirus genome downstream from the polyhedrin promoter and expressed in Sf-9 insect cells as previously described (7).The herpes simplex virus type 1 (HSV-1) DNA polymerase is a stable heterodimer composed of a large catalytic subunit (pol) and a processivity factor (UL42), both of which are essential for viral DNA synthesis in vivo and for long-chain DNA synthesis in vitro (for a review, see reference 6).

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However, an exonuclease-deficient (exo pol, the rate was slower than that for gap-filling synthesis.However, the highly conserved polymerase from another human herpesvirus, Epstein-Barr virus, was reported to possess strong strand displacement activity (31).In this report, we describe a sensitive assay to characterize the ability of wild-type and exo-deficient HSV-1 pol and pol/UL42 to engage in strand displacement synthesis.(A) A 70-mer circularized template was annealed to a 5′ end-labeled 50-mer primer strand.Extension across the ss gap would yield a reaction product 70 nt in length, while strand displacement would be characterized by products of 70 nt in length.

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