Summary
The acyclic nucleoside 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) is a potent inhibitor of human cytomegalovirus in vitro and in vivo. In order to investigate the phosphorylation of DHPG to the monophosphate and identify the enzyme responsible, attempts were made to isolate DHPG kinase from calf thymus and from human cytomegalovirus-infected lung cells. From calf thymus, a mitochondrial deoxyguanosine kinase was partially purified which co-migrated with DHPG phosphorylating activity on DEAE-cellulose, and had the same mobility by electrophoresis. DHPG triphosphate and DHPG kinase were elevated in cytomegalovirus-infected cells, but not enough enzyme activity was recovered to identify the kinase. However, DHPG was found to inhibit a cytosol deoxyguanosine kinase induced in these infected cells. The role of mitochondrial and cytosol deoxyguanosine kinases is discussed relative to the anti-cytomegalovirus activity of DHPG.
Similar content being viewed by others
References
Smith KO, Galloway KS, Kendell WL, Ogilvie KK, Radatus BK: A new nucleoside analog, 9-[[2-hydroxy-l-(hydroxymethyl)ethoxyjmethyljguanine, highly active in vitro against herpes simplex virus types 1 and 2. Antimicrob Agents Chemother 22:55–61, 1982.
Since DF, Martin JC, Verheyden JHP, Matthews TR: Antiherpesvirus activity of the acyclic nucleoside 9-(1,3-dihydroxy-2-propoxymethyl)guanine. Antimicrob Agents Chemother 23:676–682, 1983.
Cheng YC, Humng E-S, Lin J-C, Mar E-C, Pagano JS, Dutschman GE, Grill SP: Unique spectrum of activity of 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine against herpesviruses in vitro and its mode of action against herpes simplex virus type 1. Proc Natl Acad Sci 80:2767–2770, 1983.
Field AK, Davies ME, DeWitt C, Perry HC, Lion R, Germershausen J, Karkas JD, Ashton WT, Johnston DBR, Tolman RL: 9-[[2-hydroxy-l-(hydroxymethyl)-ethoxyjmethyljguanine: a selective inhibitor of herpes group virus replication. Proc Natl Acad Sci 80:4139–4143, 1983.
Tocci MJ, Livelli TJ, Perry HC, Crumpacker CS, Field AK: Effects of the nucleoside analog 2′-nor-2′-deoxyguanosine on human cytomegalovirus replication. Antimicrob Agents Chemother 25:247–252, 1984.
Mar E-C, Cheng YC, Huang E-C: Effect of 9-(1,3-dihydroxy-2-propoxymethyl)-guanine on human cytomegalovirus replication in vitro. Antimicrob Agents Chemother 24:518–521, 1984.
Mar E-C, Chiou J-F, Cheng YC, Huang E-S: Inhibition of cellular DNA polymerase a and human cytomegalovirusinduced DNA polymerase by the triphosphates of 9-(2-hydroxyethoxymethyl)guanine and 9-(1,3-dihydroxy-2-propoxymethyl)guanine. J Virol 53:776–780, 1985.
Boehme RE: Phosphorylation of the antiviral precursor 9-(1,3-dihydroxy-2-propoxymethyl)guanine monophosphate by guanylate kinase isozymes. J Biol Chem 259: 12346–12349, 1984.
Smee DF, Boehme R, Chernow M, Binko BP, Matthews TR: Intracellular metabolism and enzymatic phosphorylation of 9-(1,3-dihydroxy-2-propoxymethyl)guanine in herpes simplex virus-infected and uninfected cells. Biochem Pharmacol 34:1049–1056, 1985.
Ikeda S, Park I, Gardner P, Ives DH: New affinity adsorbents containing deoxycytidine, deoxyadenosine, or deoxyguanosine and their interactions with deoxynueleosidemetabolizing enzymes. Biochemistry 23:1914–1921, 1984.
Chen Y-C: Thymidine kinase from blast cells of myclocytic leukemia. Methods Enzymol 51:365–371, 1978.
Fyfe JA, Keller PM, Furman PA, Miller RL, Elion GB: Thymidine kinase from herpes simplex virus phosphorylates the new antiviral compound, 9-(2-hydroxyethoxymethyl)guanine. J Biol Chem 253:8723–8727, 1978.
Meijer H, Bruggeman CA, Dormans PHJ, van Boven CPA: Human cytomegalovirus induces a cellular deoxyguanosine kinase, also interacting with acyclovir. FEMS Microbiol Lett 25:283–287, 1985.
Kit S, Leung W-C, Trkula D: Properties of mitochondrial thymidine kinases of parental and enzyme-deficient HeLa cells. Arch Biochem Biophys 158:503–513, 1973.
Durham JP, Ives DH: Deoxycytidine kinase. 11. Purification and general properties of the calf thymus enzyme. J Biol Chem 245:2276–2284, 1970.
Gower WR, Jr, Carr MC, Ives DH: Deoxyguanosine kinase. Distinct molecular forms in mitochondria and cytosol. J Biol Chem 254:2180–2183, 1979.
Furman PA, DeMiranda P, St Clair MH, Elion GB: Metabolism of acyclovir in virus-infected and uninfected cells. Antimicrob Agents Chemother 20:518–524, 1981.
Meijer H, Bruggeman CA, Dormans PHJ, van Boven CPA: Rat cytomegalovirus induces cellular purine and pyrimidine nucleoside kinases in rat embryo fibroblasts and TK- Rat-2 cells. Correlations with the antiviral activity of acyclovir. Arch Virol 83:181–194, 1985.
Lewis RA, Watkins L, St.Jeor S: Enhancement of deoxyguanosine kinase activity in human lung fibroblast cells infected with human cytomegalovirus. Molec Cellular Biochem 65:67–71, 1985.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Smee, D.F. Interaction of 9-(1,3-dihydroxy-2-propoxymethyl)guanine with cytosol and mitochondrial deoxyguanosine kinases: possible role in anti-cytomegalovirus activity. Mol Cell Biochem 69, 75–81 (1985). https://doi.org/10.1007/BF00225929
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00225929