Structure-function analysis of the herpes simplex virus type 1 UL12 gene: correlation of deoxyribonuclease activity in vitro with replication function

Virology
J O HendersonD S Parris

Abstract

Although the product of the UL12 gene of herpes simplex virus type 1 (HSV-1) has been shown to possess both exonuclease and endonuclease activities in vitro, and deletion of most of the gene within the viral genome results in inefficient production and maturation of infectious virions, the function of the deoxyribonuclease (DNase) activity per se in virus replication remains unclear. In order to correlate the in vitro and in vivo activities of the protein encoded by UL12, mutant proteins were tested for nuclease activity in vitro by a novel hypersensitivity cleavage assay and for their ability to complement the replication of a DNase null mutant, AN-1. Rabbit reticulocyte lysates programmed with wild-type UL12 RNA cleaved at the same sites cleaved by purified HSV-1 DNase, but distinct from those cleaved by DNase 1 or micrococcal nuclease. All mutants which lacked DNase activity in vitro also failed to complement the replication of AN-1 in nonpermissive cells. Likewise, all mutants which contained HSV-1 DNase activity, as detected by the hypersensitivity cleavage assay, were capable of complementing the replication of the DNase null mutant, though to varying extents. Of particular note was the d1-126 mutant protein, which, despi...Continue Reading

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Citations

Sep 9, 2011·The Journal of General Virology·Alison L KuchtaMichael A McVoy
Dec 17, 2014·Antiviral Research·Zohaib AlamMichael A McVoy
Dec 29, 2013·Journal of Virology·Brett A DuguayJames R Smiley
Jul 6, 2014·Journal of Virology·Hikaru FujiiYasushi Kawaguchi
Dec 31, 2020·Viruses·Christos DogrammatzisMaria Kalamvoki

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