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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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