Methylation of HSV-1 DNA as a mechanism of viral inhibition: Studies of an analogue of methyldeoxycytidine: Trifluoromethyldeoxycytidine (F3mdCyd)

Janet Barletta, Sheldon B. Greer

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Although several hypomethylating agents such as 5-azadeoxycytidine and 5-fluorodeoxycytidine have been shown to activate transcription after incorporation into viral or cellular DNA, agents which selectively affect the methylation status of virus-infected cells have not been described. Studies on the antiviral effect of the methyldeoxycytidine (mdCyd) analogue trifluoromethyldeoxycytidine (F3mdCyd) showed significant antiviral activity against herpes simplex virus type 1 (HSV-1). This analogue of both dCyd and dThd is selectively incorporated into the DNA of herpesvirus infected cells due to the unique specificity of the herpesvirus thymidine kinase (TK) because the HSV-1 TK is both a dCyd and dThd kinase. In contrast, the deoxycytidine kinase of uninfected cells preferentially phosphorylates dCyd and has a poor affinity for F3mdCyd. F3mdCyd hemisubstituted M13 DNA displayed the same properties as mdCyd-substituted M13 DNA with respect to cleavage by restriction enzymes, and acted as an efficient template for eukaryotic DNA methyltransferase (S-adenosyl-l-methionine DNA (cytosine-5) methyltransferase: EC 2.1.1.37). Using the persistently infected CEM cell model system, the extent of DNA methylation was shown to increase in a dose-related manner when HSV-1-infected CEM cells were treated with increasing concentrations of F3mdCyd. Higher levels of methylation correlated with significant decreases in HSV-1 titers. Isoschizomer analyses followed by Southern blotting and hybridization with genomic HSV-1 DNA showed that DNA from HSV-1-infected, analogue-treated Vero cells was resistant to cleavage by restriction enzymes at a time when productive virus was not present in culture. We infer from these results that the methylation-like properties of the incorporated F3mdCyd occur concomitantly with, and appear to be involved in, the mechanisms of the analogue's antiviral effect towards HSV-1.

Original languageEnglish
Pages (from-to)1-25
Number of pages25
JournalAntiviral Research
Volume18
Issue number1
DOIs
StatePublished - Jan 1 1992
Externally publishedYes

Fingerprint

Human Herpesvirus 1
Methylation
DNA
Antiviral Agents
Thymidine Kinase
Herpesviridae
decitabine
DNA (Cytosine-5-)-Methyltransferase
Deoxycytidine Kinase
Viruses
Nucleic Acid Hybridization
Vero Cells
Methyltransferases
DNA Methylation
Enzymes
Southern Blotting
Viral Load
Methionine
Phosphotransferases

Keywords

  • HSV-1
  • Methylation
  • Trifluoromethyldeoxycytidine

ASJC Scopus subject areas

  • Virology
  • Pharmacology

Cite this

Methylation of HSV-1 DNA as a mechanism of viral inhibition : Studies of an analogue of methyldeoxycytidine: Trifluoromethyldeoxycytidine (F3mdCyd). / Barletta, Janet; Greer, Sheldon B.

In: Antiviral Research, Vol. 18, No. 1, 01.01.1992, p. 1-25.

Research output: Contribution to journalArticle

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