title: Phosphorylation of the carbocyclic analog of 2'-deoxyguanosine in cells infected with herpes viruses.
abstract: The carbocyclic analog of 2'-deoxyguanosine [(+-)-2-amino-1,9-dihydro-9-[(1 alpha,3 beta,4 alpha)-3-hydroxy-4-(hydroxymethyl)cyclopentyl]-6H-purine-6-one] (2'-CDG) is highly active in cell culture against strains S148 and E377 of herpes simplex virus type 1 (HSV-1), both of which code for thymidine kinase, and much less active against strain BW10168 which is deficient in this enzyme activity. Antiviral activity is associated primarily with the D-enantiomer; the L-enantiomer has much lower but significant activity. The metabolism of racemic 2'-CDG and its D- and L-enantiomers was studied in uninfected HEp-2 cells and in HEp-2 cells infected with the S148 or BW10168 strains of HSV-1. Nucleotides were separated by HPLC, and their elution was monitored by spectrophotometry. The chromatograms of extracts of cells infected with the S148 strain and treated with (+/-)-2'-CDG or D-2'-CDG included a new peak which appeared in the triphosphate region. This peak, the area of which exceeded that of the GTP peak, was shown to be due to the triphosphate of 2'-CDG. The new peak was not observed by HPLC of extracts of uninfected cells treated with (+/-)-2'-CDG or either of its enantiomers, cells infected with the S148 strain and treated with L-2'-CDG, or cells infected with the BW10168 strain and treated with (+/-)-2'-CDG or either of its enantiomers. The results were similar when these studies were performed with uninfected Vero cells and with Vero cells infected with strain S148 of HSV-1. In experiments with D-[8-3H]-2'-CDG, small amounts of phosphates of 2'-CDG could also be detected in uninfected HEp-2 cells and in cells infected with the BW10168 strain of HSV-1. Thus, 2'-CDG apparently is a good substrate for the virus-coded kinase and a very poor substrate for cellular phosphorylating enzymes. The selective phosphorylation of 2'-CDG by the virus-specific kinase presumably is critical for its antiviral activity as it is for that of acyclovir and other acyclic derivatives of guanine.
authors.affiliation: Kettering-Meyer Laboratory, Southern Research Institute, Birmingham, AL 35255.
authors.forename: L L
authors.initials: LL
authors.lastname: Bennett
authors.forename: Y F
authors.initials: YF
authors.lastname: Shealy
authors.forename: P W
authors.initials: PW
authors.lastname: Allan
authors.forename: L M
authors.initials: LM
authors.lastname: Rose
authors.forename: W M
authors.initials: WM
authors.lastname: Shannon
authors.forename: G
authors.initials: G
authors.lastname: Arnett
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D006561:Herpes Simplex / Q000378:metabolism; D009154:Mutation; D009711:Nucleotides / Q000302:isolation & purification; D010766:Phosphorylation; D018139:Simplexvirus / Q000201:enzymology* / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D009711:Nucleotides; C061610:2-amino-1,9-dihydro-9-(3-hydroxy-4-(hydroxymethyl)cyclopentyl)-6H-purine-6-one; D013937:Thymidine Kinase; D003849:Deoxyguanosine