Immunological evidence for a specific DNA polymerase produced after infection by herpes simplex virus

115

SUMMARYRibonucleotide reductase, an enzyme found in all prokaryotic and eukaryotic cells that synthesize DNA, is induced by herpes simplex virus (HSV). In this study the effect of anti-HSV antiserum on the induced ribonucleotide reductase has been examined and the ability of different temperature-sensitive (ts) mutants of HSV-1 to induce the enzyme has been investigated. The HSV-l-induced ribonucleotide reductase was inhibited by antiserum raised against infected cell lysates but not by preimmune serum.The wild-type (ts +) virus induced similar levels of ribonucleotide reductase at 31 °C and 38-5 °C (the permissive and non-permissive temperatures respectively for the ts mutants). All ts mutants induced approximately wild-type levels of the enzyme at 31 °C. At 38.5 °C, two of the four ts mutants studied also induced wild-type levels of enzyme but ts G failed to induce any activity while ts K induced variable but low levels. The enzyme activity induced by ts G at 31 °C was thermolabile both in vivo and in vitro. These results provide the first strong evidence that the induced ribonucleotide reductase activity is at least partially virus-coded.

supporting

confidence: 61%

Ribonucleotide Reductase Induced by Herpes Simplex Virus has a Virus-specified Constituent

Dutia

1983

115

“…Multiple virus specific antigens have been recognized in cells infected with herpes simplex virus (Tokumaru, 1965;Watson et al I966;Honess et al 1974). Some of these represent antigenic activities which are also present as components of the virus particle (e.g.…”

Section: Introductionmentioning

confidence: 99%

“…As well as containing type specific and cross neutralizing antibody against infectious virus particles (Sire & Watson, 1973) these sera also contained antibodies to a number of virus specific enzyme activities (Keir et al I966;Klemperer et al 1967;Morrison & Keir, 1967;Buchan & Watson, 1969;Buchan et al I97oa, b).…”

Section: Introductionmentioning

confidence: 99%

Herpes Simplex Virus-specific Polypeptides Studied by Polyacrylamide Gel Electrophoresis of Immune Precipitates

Honess¹,

Watson²

1974

SUMMARYAntisera raised in rabbits against RK13 cells infected with herpes simplex virus type I were capable of specifically precipitating proteins synthesized after infection of BHK-2I cells with the virus. Analysis of these immune precipitates by polyacrylamide gel electrophoresis demonstrated ~> 15 polypeptides with mol. wt. from 25 to IOOOOO. A number of these polypeptides were not detected in purified preparations of virus particles.Precipitates formed with two 'monoprecipitin' antisera were also analysed. Antiserum to the structural antigen Band II precipitated a major polypeptide of mol. wt. 470o0, which was glycosylated, and corresponded in mobility to a minor component polypeptide of the herpes virus particle. The other monoprecipitin antiserum, to the herpes-specified thymidine kinase, precipitated a polypeptide with a tool. wt. of 44000. The thymidine kinase polypeptide was not glycosylated.

“…Cell infection and crude enzyme extract preparation were as given for exonuclease activity. The assay was essentially as described by Keir et al (1966), except that samples were taken at 30 and 60 min.…”

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Revertants from Fourteen Herpes Simplex Virus Type 1 (Strain 17) Temperature-sensitive Mutants

Dargan

Subak‐Sharpe

1984

SUMMARYThe isolation of spontaneous non-temperature-sensitive revertants of herpes simplex virus type 1 (HSV-1) (strain 17) ts mutants, following a single round of selection at nonpermissive temperature, is reported. The distinction between total and incomplete reversion was assessed by the wild-type performance of non-ts revertants in three aspects of HSV infection: (i) virus growth, (ii) induction of HSV-specified enzyme activities (where appropriate) and (iii) the gel profile of infected cell polypeptides. Wild-type behaviour was taken to indicate total reversion, while less than wild-type performance in any test indicated incomplete reversion. Only five of 19 HSV-1 mutants (ts L, U, X, R and G syn) examined for reversion of the ts lesion failed to yield non-ts revertants; these mutants may well contain multiple mutations. Total reversion was obtained with mutants ts