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Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293.

Publication ,  Journal Article
Upton, DC; Wang, X; Blans, P; Perrino, FW; Fishbein, JC; Akman, SA
Published in: Chemical research in toxicology
July 2006

N(2)-Ethyldeoxyguanosine (N(2)-ethyldGuo) is a DNA adduct formed by reaction of the exocyclic amine of dGuo with the ethanol metabolite acetaldehyde. Because ethanol is a human carcinogen, we assessed the biological consequences of replication of template N(2)-ethyldGuo, in comparison to the well-studied adduct O(6)-ethyldeoxyguanosine (O(6)-ethyldGuo). Single chemically synthesized N(2)-ethyldGuo or O(6)-ethyldGuo adducts were placed site specifically in the suppressor tRNA gene of the mutation reporting shuttle plasmid pLSX. N(2)-EthyldGuo and O(6)-ethyldGuo were both minimally mutagenic in double-stranded pLSX replicated in human 293 cells; however, the placement of deoxyuridines on the complementary strand at 5'- and 3'-positions flanking the adduct resulted in 5- and 22-fold enhancements of the N(2)-ethyldGuo- and O(6)-ethyldGuo-induced mutant fractions, respectively. The fold increase in the N(2)-ethyldGuo-induced mutant fraction in deoxyuridine-containing plasmids was similar after replication in 293T cells, a mismatch repair deficient variant of 293 cells, indicating that postreplication mismatch repair has little role in modulating N(2)-ethyldGuo-mediated mutagenesis. The mutation spectrum generated by N(2)-ethyldGuo consisted primarily of single base deletions and adduct site-targeted transversions, in contrast to the exclusive production of adduct site-targeted transitions by O(6)-ethyldGuo. The yield of progeny plasmids after replication in 293 cells was reduced by the presence of N(2)-ethyldGuo in parental plasmids with or without deoxyuridine to 39 or 19%, respectively. Taken together, these data indicate that N(2)-ethyldGuo in DNA exerts its principal biological activity by blocking translesion DNA synthesis in human cells, resulting in either failure of replication or frameshift deletion mutations.

Published In

Chemical research in toxicology

DOI

EISSN

1520-5010

ISSN

0893-228X

Publication Date

July 2006

Volume

19

Issue

7

Start / End Page

960 / 967

Related Subject Headings

  • Toxicology
  • Plasmids
  • Mutagenicity Tests
  • Molecular Sequence Data
  • Kidney
  • Humans
  • Genetic Vectors
  • Frameshift Mutation
  • Deoxyguanosine
  • DNA Replication
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Upton, D. C., Wang, X., Blans, P., Perrino, F. W., Fishbein, J. C., & Akman, S. A. (2006). Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293. Chemical Research in Toxicology, 19(7), 960–967. https://doi.org/10.1021/tx060084a
Upton, Dana C., Xueying Wang, Patrick Blans, Fred W. Perrino, James C. Fishbein, and Steven A. Akman. “Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293.Chemical Research in Toxicology 19, no. 7 (July 2006): 960–67. https://doi.org/10.1021/tx060084a.
Upton DC, Wang X, Blans P, Perrino FW, Fishbein JC, Akman SA. Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293. Chemical research in toxicology. 2006 Jul;19(7):960–7.
Upton, Dana C., et al. “Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293.Chemical Research in Toxicology, vol. 19, no. 7, July 2006, pp. 960–67. Epmc, doi:10.1021/tx060084a.
Upton DC, Wang X, Blans P, Perrino FW, Fishbein JC, Akman SA. Replication of N2-ethyldeoxyguanosine DNA adducts in the human embryonic kidney cell line 293. Chemical research in toxicology. 2006 Jul;19(7):960–967.
Journal cover image

Published In

Chemical research in toxicology

DOI

EISSN

1520-5010

ISSN

0893-228X

Publication Date

July 2006

Volume

19

Issue

7

Start / End Page

960 / 967

Related Subject Headings

  • Toxicology
  • Plasmids
  • Mutagenicity Tests
  • Molecular Sequence Data
  • Kidney
  • Humans
  • Genetic Vectors
  • Frameshift Mutation
  • Deoxyguanosine
  • DNA Replication