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Regulation of herpesvirus thymidine kinase activity in lm thymidine kinase negative cells transformed by uv light irradiated herpes simplex virus


, : Regulation of herpesvirus thymidine kinase activity in lm thymidine kinase negative cells transformed by uv light irradiated herpes simplex virus. Virology 76(1): 331-340

Thymidine kinase (TK) activity of herpes simplex virus-transformed cells [mouse fibroblast] [LM(TK-)/HSV-1 and LM(TK-)/HSV-2] was stimulated 6-9 h after the cells were infected with TK-negative mutants of either type 1 or type 2 herpes simplex virus (HSV-1 TK-, HSV-2 TK-) or with TK-negative mutants of marmoset herpesvirus (MarHV TK-) and pseudorabies virus (PRV TK-). Inhibitors of DNA synthesis did not prevent this stimulation, but inhibitors of RNA and protein synthesis did. Equine herpesvirus (EHV-1) and TK-negative mutants of vaccinia virus did not enhance the TK activity of the transformed cells, showing that stimulation was not a consequence merely of nonspecific virus infection. HSV-1 B2015, a mutant deficient in inducing TK activity in productively infected LM(TK-) cells at 37.degree. C stimulated HSV-transformed cell TK at this temperature, suggesting that the gene which controls TK turn-on probably functions normally in mutant B2015. Enhancement by TK-negative herpesviruses can apparently occur regardless of whether the superinfecting herpesvirus produces an inactive TK polypeptide or none at all. Autoradiographic and enzyme studies on a subline of LM(TK-)/HSV-1 cells grown in nonselective medium without HAT (hypoxanthine, aminopterin and thymidine) demonstrated that essentially all of the cells contained significant TK activity. The TK activity of this subline and of clonal sublines isolated in nonselective medium or in counterselective medium with bromodeoxyuridine (BrUdR) was enhanced by HSV-1 TK- infection, supporting the suggestion by Davidson and co-workers that the HSV TK gene was retained in most transformed cells grown in nonselective medium. Immunological studies, using type-specific immunoglobulin (IgG) fractions prepared from sera of rabbits that were immunized with HSV-1 TK or HSV-2 TK, demonstrated that the enhanced TK activity of HSV-1-transformed cells was inhibited by anti-HSV-1 IgG and that of HSV-2-transformed cells was inhibited by anti-HSV-2 IgG, regardless of whether the transformed cells were superinfected with HSV-1 TK-, HSV-2 TK- or MarHV TK-. Neither antiserum inhibited the TK activity induced by wild-type MarHV in LM(TK-) cells, indicating that the TK activity stimulated by TK-negative herpesvirus infections of HSV-transformed cells had the antigenic specificity of the resident virus TK.

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Related references

Kit, S.; Dubbs, D.R., 1977: Regulation of herpesvirus thymidine kinase activity in LM(TK) cells transformed by ultraviolet light-irradiated herpes simplex virus. Virology 76(1): 331-340

Kit S.; Dubbs D.R.; Schaffer P.A., 1977: Thymidine kinase activity of herpes simplex virus transformed cells super infected with thymidine kinase negative temperature sensitive herpes simplex virus mutants. Abstracts of the Annual Meeting of the American Society for Microbiology 77: 313

Kit, S.; Dubbs, D.R.; Schaffer, P.A., 1978: Thymidine kinase activity of biochemically transformed mouse cells after super infection by thymidine kinase negative temperature sensitive herpes simplex virus mutants. Previous studies showed that thymidine kinase (TK)-negative mutants isolated from HSV[herpes simplex virus]-1 clone 101, HSV-2 (333), marmoset herpesvirus and pseudorabies virus enhanced the resident TK [thymidine kinase] activity of biochemically...

Kit, S.; Otsuka, H.; Qavi, H.; Hazen, M., 1981: Herpes simplex virus thymidine kinase activity of thymidine kinase-deficient Escherichia coli K-12 mutant transformed by hybrid plasmids. A hybrid plasmid (pAGO) that contains the herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene in the form of a 2-kilobase-pair (kbp) Pvu II fragment inserted at the Pvu II site of plasmid pBR322 was used to transform TK- Escherichia col...

Suzutani, T.; Koyano, S.; Takada, M.; Yoshida, I.; Azuma, M., 1995: Analysis of the relationship between cellular thymidine kinase activity and virulence of thymidine kinase-negative herpes simplex virus types 1 and 2. The virulence of thymidine kinase-negative herpes simplex virus type 1 (HSV-1; VRTK- strain) and type 2 (HSV-2; UWTK- strain) was studied in comparison with that of their parental strains (VR-3 and UW-268, respectively) in an encephalitis model of...

Bastow, K.F.; Darby, G.; Wildy, P.; Minson, A.C., 1980: Properties of cells carrying the herpes simplex virus type 2 thymidine kinase gene: mechanisms of reversion to a thymidine kinase-negative phenotype. We have isolated cells with a thymidine kinase-negative (tk(-)) phenotype from cells which carry the herpes simplex virus type 2 tk gene by selection in 5-bromodeoxyuridine or 9-(2-hydroxyethoxymethyl)guanine. Both selection routines generated rev...

Sheflin L.G.; Chadha K.C.; Zeigel R.F.; Hughes R.G.Jr, 1977: Transformation of l thymidine kinase negative cells to the thymidine kinase positive phenotype is not effected by noninfectious or defective interfering particles of herpes simplex virus type 1. Abstracts of the Annual Meeting of the American Society for Microbiology 77: 286

Tenser R.B.; Edris W.A., 1986: Thymidine kinase activity in herpes simplex virus type 1 recombinants that carry insertions affecting regulation of the thymidine kinase gene. Determinations of the possible importance of herpes simplex virus type 1 (HSV) thymidine kinase (TK) expression in the pathogenesis of viral latency depend in part on the use of defined mutants. In a recent study by A.E. Sears, B. Meignier, and B....

Chen, M.S.; Summers, W.P.; Walker, J.; Summers, W.C.; Prusoff, W.H., 1979: Characterization of pyrimidine deoxyribonucleoside kinase (thymidine kinase) and thymidylate kinase as a multifunctional enzyme in cells transformed by herpes simplex virus type 1 and in cells infected with mutant strains of herpes simplex virus. Pyrimidine deoxyribonucleoside kinase (thymidine kinase [TK]) was purified from two herpes simplex virus type 1 (HVS-1)-transformed TK-deficient mouse (LMTK-) cell lines and from LMTK- cells infected with HSV-1 mutant viruses coding for variant TK...

Chen M.S.; Summers W.P.; Walker J.; Summers W.C.; Prusoff W.H., 1979: Characterization of pyrimidine deoxy ribo nucleoside kinase thymidine kinase and thymidylate kinase as a multi functional enzyme in cells transformed by herpes simplex virus type 1 and in cells infected with mutant strains of herpes simplex virus. Pyrimidine deoxyribonucleoside kinase (thymidine kinase [TK]) was purified from 2 herpes simplex virus type 1 (HSV-1)-transformed TK-deficient mouse (LMTK-) cell lines and from LMTK- cells infected with HSV-1 mutant viruses coding for variant TK e...