Induction of thymidine kinase by vaccinia-infected mouse fibroblasts

Kit, Saul; Piekarski, L; Dubbs, D. R.

doi:10.1016/s0022-2836(63)80078-9

Cited by 112 publications

(31 citation statements)

References 20 publications

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“…Involvement of thymidine kinase activity during infection of cells with DNA viruses and DNA tumor viruses (25)(26)(27) and during induction with IdUrd of the Epstein-Barr virus in Burkitt lymphoblastoid cells (28,29) has previously been shown.…”

Section: Inhibition Of Virusmentioning

confidence: 90%

Induction of Endogenous Virus and of Thymidine Kinase by Bromodeoxyuridine in Cell Cultures Transformed by Friend Virus

Ostertag

Roesler

Krieg

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

106

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Thymidine kinase positive (TK+) N type cell lines that had been transformed by spleen focus-forming virus were established by transformation with NB tropic Friend virus complex. Thymidine kinase deficient (TK-) cell clones were isolated. Some of these cell clones release 1000-to 100,000-fold reduced amounts of Friend virus complex as compared to the TK+ parental cell clone. TK-clones were grown in medium without BrdUrd. Some of these TK-clones can be induced to release endogenous helper virus and transforming spleen focus-forming virus on reexposure to 10-10-4 M BrdUrd. BrdUrd inhibits the replication of C-type tumor viruses (1), whereas in some instances BrdUrd or IdUrd can induce endogenous C-type virus (2-4). In order to study the action of BrdUrd on virus replication and its role in the induction of endogenous virus we have used Friend virus (FV) infected and transformed DBA-2 mouse spleen cells in culture. The FV complex consists of two components, the lymphatic leukemia helper virus (LLV-F) and the replication-defective erythroid cell-transforming spleen focus-forming virus (SFFV-F) (5, 6). The RNA of the two virus types is presumably different in size and structure (7). We have established various cell lines transformed by the SFFV-F (8, 9) which, after another thymidine analogue, can be phosphorylated to azidothymidine triphosphate. However, the presence of an No group at the 3' terminal of the deoxysugar interferes with esterification, so presumably azidothymidine can only be added terminally to the growing DNA strand. Both azidothymidine as well as BrdUrd can be used to decrease the virus titer of BrdUrd-sensitive erythroleukemic cells. We have isolated several BrdUrd-resistant clones by treatment of sensitive cells with high doses of BrdUrd (11) and have shown that some of these clones have either decreased or barely detectable thymidine kinase (ATP: thymidine 5'-phosphotransferase, EC 2.7.1.75) activity (11). Furthermore, they have a 108-to 104-fold decreased virus titer. The decrease in C-type virus release can also be shown by electron microscopy.A temporary thymidine kinase activation is observed if BrdUrd is added to some virus negative lines. The same cells release large numbers of endogenous C-type and spleen focusforming virus on exposure to BrdUrd but not to azidothymidine. The RNA tumor virus which is induced by BrdUrd has mainly N-type host range properties unlike the original NB type FV complex which has been used for the transformation of our erythroid cell lines. Thymidine kinase induction and virus induction are always correlated. We have independent evidence (Ostertag, Crozier, and Swetly, unpublished)

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Section: Inhibition Of Virusmentioning

confidence: 90%

Induction of Endogenous Virus and of Thymidine Kinase by Bromodeoxyuridine in Cell Cultures Transformed by Friend Virus

Ostertag

Roesler

Krieg

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

106

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“…Veto cells (CCL-81) were obtained from the American Type Culture Collection (ATCC) and grown in EMEM supplemented with 5 ~ FCS and antibiotics. LM(TK-) cells (Kit et al, 1963) were kindly provided by Dr S. Kit (Baylor College of Medicine, Houston, Tx., U.S.A.) and grown as described for rabbit skin cells.…”

Section: Methodsmentioning

confidence: 99%

Biological Basis for Virulence of Three Strains of Herpes Simplex Virus Type 1

Sedarati¹,

Stevens²

1987

Journal of General Virology

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SUMMARYHerpes simplex virus type 1 (HSV-1) strains F, HF and HFEM were studied with respect to pathogenicity in mice and growth characteristics in vivo and in vitro compared to the neurovirulent HSV-I strains 17 syn + and KOS. All three viruses demonstrated reduced virulence in mouse brains and were completely avirulent after footpad inoculation. They were shown to express high levels of thymidine kinase activity. Investigations concerning the virulence phenotype indicated that the defect(s) in strains F, HF and HFEM related to general replication deficiency in mouse cells. It was also shown that although the replication restriction observed for strains F and HF was specific for murine cells, strain HFEM did not replicate well in any cell type tested. Additional studies indicated that the avirulence phenotype which followed peripheral inoculation was related to different genotypes, since strain F complemented HF and HFEM and, as expected, the latter two agents did not complement each other. All three agents were found to complement the non-neuroinvasive strain KOS. Finally, the data also show that two herpesviruses which are highly restricted in murine cells (e.g. strains F and HF) can still interact in the animal and produce a lethal infection.

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“…While herpesviruses and orthopoxviruses both express proteins with TK activity (39,40), the enzymes are distinct in several fundamental respects, including molecular weight, quaternary structure, and substrate specificity. Herpesvirus enzymes belong to the type I family of TKs, whereas the VV TK is a type II enzyme (5).…”

mentioning

confidence: 99%

Selective Phosphorylation of Antiviral Drugs by Vaccinia Virus Thymidine Kinase

Prichard¹,

Keith²,

Johnson³

et al. 2007

Antimicrob Agents Chemother

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The antiviral activity of a new series of thymidine analogs was determined against vaccinia virus (VV), cowpox virus (CV), herpes simplex virus, and varicella-zoster virus. Several compounds were identified that had good activity against each of the viruses, including a set of novel 5-substituted deoxyuridine analogs. To investigate the possibility that these drugs might be phosphorylated preferentially by the viral thymidine kinase (TK) homologs, the antiviral activities of these compounds were also assessed using TK-deficient strains of some of these viruses. Some of these compounds were shown to be much less effective in the absence of a functional TK gene in CV, which was unexpected given the high degree of amino acid identity between this enzyme and its cellular homolog. This unanticipated result suggested that the CV TK was important in the mechanism of action of these compounds and also that it might phosphorylate a wider variety of substrates than other type II enzymes. To confirm these data, we expressed the VV TK and human TK1 in bacteria and isolated the purified enzymes. Enzymatic assays demonstrated that the viral TK could efficiently phosphorylate many of these compounds, whereas most of the compounds were very poor substrates for the cellular kinase, TK1. Thus, the specific phosphorylation of these compounds by the viral kinase may be sufficient to explain the TK dependence. This unexpected result suggests that selective phosphorylation by the viral kinase may be a promising new approach in the discovery of highly selective inhibitors of orthopoxvirus replication.

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Induction of thymidine kinase by vaccinia-infected mouse fibroblasts

Cited by 112 publications

References 20 publications

Induction of Endogenous Virus and of Thymidine Kinase by Bromodeoxyuridine in Cell Cultures Transformed by Friend Virus

Induction of Endogenous Virus and of Thymidine Kinase by Bromodeoxyuridine in Cell Cultures Transformed by Friend Virus

Biological Basis for Virulence of Three Strains of Herpes Simplex Virus Type 1

Selective Phosphorylation of Antiviral Drugs by Vaccinia Virus Thymidine Kinase

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