Analysis of mutations in the Tk gene of Tk(+/-) mice treated as neonates with 3'-azido-3'-deoxythymidine (AZT).
Mother-to-child transmission of the human immunodeficiency virus (HIV) is reduced by perinatal treatment with the antiretroviral nucleoside analogue 3'-azido-3'-deoxythymidine (AZT, zidovudine). AZT, however, is genotoxic and carcinogenic in mice when administered either transplacentally or neonatally, suggesting a possible cancer risk for children later in life. In a previous study we found that treating B6C3F1/Tk(+/-) mice on postnatal days 1 through 8 with intraperitoneal injections of 200 mg AZT per kg body weight per day significantly increased spleen lymphocyte mutant frequencies in the autosomal Tk gene. Allele-specific PCR of Tk mutants from treated mice indicated that 61% had lost the Tk(+) allele (loss of heterozygosity; LOH), compared with 35% of Tk mutants from control mice, a difference that was significant. In the present study, Tk mutant lymphocyte clones were analyzed further using polymorphic microsatellite markers that flank the Tk gene along the length of mouse chromosome 11. The analysis indicated that allele-loss mutations in control mice were due to either total chromosome loss, mitotic recombination, or both. The pattern of marker loss in mutants from AZT-treated mice differed significantly from the control mice and was consistent with chromosome loss, mitotic recombination, interstitial deletion, gene conversion, and an unusual discontinuous LOH. The results indicate that AZT induced a unique pattern of mutations in the Tk gene of mice and that the major mechanisms of mutation by AZT involved deletion and recombination.
Mittelstaedt
Roberta A
RA
Division of Genetic and Reproductive Toxicology, US FDA/National Center for Toxicological Research, 3900 NCTR Road, Jefferson, AR 72079, USA. rmittelstaedt@nctr.fda.gov
Von Tungeln
Linda S
LS
Shaddock
Joseph G
JG
Dobrovolsky
Vasily N
VN
Beland
Frederick A
FA
Heflich
Robert H
RH
D000483:Alleles; D000818:Animals; D000831:Animals, Newborn; D019380:Anti-HIV Agents; D002875:Chromosomes / Q000187:drug effects; D002999:Clone Cells; D004252:DNA Mutational Analysis; D017353:Gene Deletion; D006579:Heterozygote; D019656:Loss of Heterozygosity / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects; D051379:Mice; D008822:Mice, Transgenic; D018895:Microsatellite Repeats / Q000187:drug effects; D009154:Mutation / Q000187:drug effects*; D011995:Recombination, Genetic / Q000187:drug effects; D013937:Thymidine Kinase / Q000235:genetics*; D015215:Zidovudine / Q000494:pharmacology*
D019380:Anti-HIV Agents; D015215:Zidovudine; D013937:Thymidine Kinase