Metabolic fate of [3H]1-beta-D-arabinofuranosyl-5-[(E)-2-bromovinyl]uracil in herpes simplex virus type 1-infected cells.
The metabolic fate of 1-beta-D-arabinofuranosyl-5-[(E)-2-bromovinyl]uracil (BV-araU) in herpes simplex virus type 1-infected cells was studied using tritium-labeled BV-araU. [3H]BV-araU was selectively taken-up by infected cells. Approximately 10% of the total uptake of [3H]BV-araU was recovered from the acid-insoluble fraction at any time post-infection. Both cellular uptake of [3H]BV-araU and its incorporation into the acid-insoluble fraction increased with increasing incubation time through 8 hr post-infection. Uptake of [3H]BV-araU and its incorporation into the acid-insoluble fraction also increased proportionally to the duration of exposure to [3H]BV-araU. An alkaline sucrose gradient sedimentation analysis revealed that the radioactive DNA obtained from cells pulse-labeled with [3H]BV-araU were small DNA fragments which remained at the top following a chasing period in isotope-free medium, whereas that pulse-labeled with [3H]thymidine was chased to a fraction of high molecular weight DNA. Nuclease P1 digestion reduced 99% of the [3H]BV-araU-labeled DNA extracted from infected cells to a low molecular weight. Following digestion of [3H]BV-araU-labeled DNA with micrococcal nuclease and spleen exonuclease, all of the radioactivity was recovered as [3H]BV-araU 3'-monophosphate. Thus, BV-araU strongly inhibits the elongation of viral DNA strands as demonstrated by the alkaline sucrose gradient sedimentation analysis, whereas at least a portion of the [3H]BV-araU is incorporated inside viral DNA strands in infected cells.
Ashida
N
N
Biology Laboratory, Yamasa Corporation, Choshi, Chiba, Japan.
Sakata
S
S
Machida
H
H
D000212:Acyclovir / Q000378:metabolism; D000998:Antiviral Agents / Q000378:metabolism* / Q000494:pharmacology; D001086:Arabinofuranosyluracil / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002851:Chromatography, High Pressure Liquid; D004279:DNA, Viral / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000032:analysis; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D000998:Antiviral Agents; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D014316:Tritium; D001086:Arabinofuranosyluracil; C031368:sorivudine; D003851:Deoxyribonucleases; D013936:Thymidine; D000212:Acyclovir