Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines

Stinchcombe, Timothy; Clough, Wendy

doi:10.1021/bi00329a034

Cited by 20 publications

(21 citation statements)

References 33 publications

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“…Virus-nonproducing, 5-bromodeoxyuridine-resistant P3HRI cells were induced to produce virus by removal of the drug, and TK activity was demonstrated in these cells (Hampar et al,I97I). The presence of virus-specific TK in EBV-infected Iymphoblastoid cells has also been demonstrated in several laboratories (Glaser et al, 1973;Kit et al, 1975;Chen et al, 1978;Graessmann eta]., 1980;Ooka & Calender, I980;Roubal & Klein, 1981;MacGabhann et al, 1984;Stinchcombe & Clough, 1985 ;de Turenne-Tessier et al, 1986). However, purification of TK protein from these cells was not successful due to its low abundance following induction of EBV-containing cells and the difficulty in separating it from cellular TK.…”

Section: Introductionmentioning

confidence: 76%

Functional analysis of C-terminal deletion mutants of Epstein--Barr virus thymidine kinase

Hsu

Liu

Chang

et al. 1996

Journal of General Virology

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Thymidine kinase fiR) activity was detected following expression of the TK gene of Epstein-Barr virus (EBV) using the pET expression plasmid and E. coli BL21(DE3)pLysS. To study the amino acid residues required at the C terminus of the EBV TK protein for enzymatic activity, a series of C-terminal deletion mutants was generated by direct truncation, linker insertion or PCR mutagenesis to create stop codons at particular sites. Deletion of nine residues from the C terminus caused a 35% reduction in TK activity, while a ten-residue deletion completely abolished the activity. A single point mutation at residue Cys570, corresponding to Cys336 of herpes simplex virus TK, did not alter the TK activity. Single amino acid changes within the last seven to ten residues also did not affect activity. The results indicate that maintenance of the conformation of the C terminus is important for enzyme activity.

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Section: Introductionmentioning

confidence: 76%

Functional analysis of C-terminal deletion mutants of Epstein--Barr virus thymidine kinase

Hsu

Liu

Chang

et al. 1996

Journal of General Virology

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“…When the drugs were removed for 25 days, the linear viral DNA reappeared, suggesting the presence of a small amount of replicating virus (28). The virus-specific TK protein is demonstrable in H1 EBV replicating cells but not demonstrable in L5 cells (31,45). The metabolism of tritiated L-IOddU in H1 and L5 cells showed no phosphorylated metabolites in L5 but showed the formation of monophosphate as well as very small amounts of the putative di-and triphosphate forms in H1.…”

Section: Discussionmentioning

confidence: 98%

Anti-Epstein-Barr Virus (EBV) Activity of β- l -5-Iododioxolane Uracil Is Dependent on EBV Thymidine Kinase

Kira

Grill

Dutschman

et al. 2000

Antimicrob Agents Chemother

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␤-L-5-Iododioxolane uracil was shown to have potent anti-Epstein-Barr virus (EBV) activity (50% effective concentration ‫؍‬ 0.03 M) with low cytotoxicity (50% cytotoxic concentration ‫؍‬ 1,000 M). It exerts its antiviral activity by suppressing replicative EBV DNA and viral protein synthesis. This compound is phosphorylated in cells where the EBV is replicating but not in cells where the EBV is latent. EBV-specific thymidine kinase could phosphorylate ␤-L-5-iododioxolane uracil to the monophosphate metabolite. The K m of ␤-L-5-iododioxolane uracil with EBV thymidine kinase was estimated to be 5.5 M, which is similar to that obtained with thymidine but about fivefold higher than that obtained with 2 fluoro-5-methyl-␤-L-arabinofuranosyl uracil, the first L-nucleoside analogue discovered to have anti-EBV activity. The relative V max is seven times higher than that of thymidine. The anti-EBV activity of ␤-L-5-iododioxolane uracil and its intracellular phosphorylation could be inhibited by 5-ethynylthymidine, a potent EBV thymidine kinase inhibitor. The present study suggests that ␤-L-5-iododioxolane uracil exerts its action after phosphorylation; therefore, EBV thymidine kinase is critical for the antiviral action of this drug. Epstein-Barr virus (EBV) is an important human pathogen.This virus has been determined to cause infectious mononucleosis, fatal acute infectious mononucleosis-X-linked lymphoproliferative syndrome, and oral hairy leukoplakia (16,17,48). EBV also has a close association with several types of malignancies, including Burkitt's lymphoma, nasopharyngeal carcinoma (NPC), Hodgkin's disease, (2, 13, 21), some T-cell lymphomas (1,26,32,35,40,58), smooth muscle cell leiomyosarcoma (23), and certain cases of gastric adenocarcinomas (38,46,53). EBV infection can enhance human immunodeficiency virus type 1 (HIV-1) replication in T cells (56), and EBV is also related to the development of lymphoma induced in AIDS patients (55). It was also reported that almost all posttransplant lymphomas appeared to be EBV genome positive, irrespective of histological appearance or clonability of the lesion (18,44,49,59). A more complex picture of EBV-associated malignancies is emerging, particularly with regard to virus-positive tumors of non-B-cell origin. It is hoped that a better understanding of EBV persistence and the part played by EBV in the oncogenic process will permit the development of new approaches aimed at the prevention and treatment of EBV-associated tumors. Therefore, it would be ultimately useful to have anti-EBV compounds that do not have serious side effects.Several compounds have shown anti-EBV activity in cell culture systems, including acyclovir (ACV), ganciclovir (DHPG), 2Ј-fluoro-5-methyl-␤-D-arabinofuranosyl uracil (D-FMAU), and (S)-1-(3-hydroxy-2-phosphonyl-methoxypropyl)cytosine (cidofivir) (6,9,26,27,36,50,52). However, the clinical application of many of these compounds is restricted by severe side effects (52). Recently, our laboratory found a new anti-EBV L-dioxolane nucleoside analogue, ␤...

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“…30 In a rate-limiting step, the TK from these viruses can convert nucleoside analogs to their monophosphate form. 30,31 Cellular enzymes then complete their conversion to biologically active triphosphates. A viral DNA polymerase preferentially incorporates the toxic metabolites into viral DNA, leading to premature termination of the nascent DNA, with resulting apoptosis of infected cells.…”

Section: Introductionmentioning

confidence: 99%

A phase 1/2 trial of arginine butyrate and ganciclovir in patients with Epstein-Barr virus–associated lymphoid malignancies

Perrine

Hermine

Small

et al. 2006

Blood

265

179

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Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines

Cited by 20 publications

References 33 publications

Functional analysis of C-terminal deletion mutants of Epstein--Barr virus thymidine kinase

Functional analysis of C-terminal deletion mutants of Epstein--Barr virus thymidine kinase

Anti-Epstein-Barr Virus (EBV) Activity of β- l -5-Iododioxolane Uracil Is Dependent on EBV Thymidine Kinase

A phase 1/2 trial of arginine butyrate and ganciclovir in patients with Epstein-Barr virus–associated lymphoid malignancies

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