Mutagenesis by acrolein-derived propanodeoxyguanosine adducts in human cells.

Abstract

Acrolein, which is widely spread in the environment and is produced by lipid peroxidation in cells, reacts with DNA to form two exocyclic 1,N2-propanodeoxyguanosine (PdG) adducts. To establish their relative contribution to the acrolein mutagenicity, the genotoxic properties of alpha-OH-PdG and gamma-OH-PdG together with their model DNA adduct, PdG, were studied in human cells. DNA adducts were incorporated site-specifically into a SV40/BK virus origin-based shuttle vector and replicated in xeroderma pigmentosum complementation group A (XPA) cells. Analysis of progeny plasmid revealed that alpha-OH-PdG and PdG strongly block DNA synthesis and that both adducts induced base substitutions with G --> T transversions predominating. Primer extension studies, catalyzed by the 3'-->5' exonuclease-deficient Klenow fragment of Escherichia coli pol I, revealed limited extension from the 3' primer termini opposite these two adducts. In contrast, gamma-OH-PdG did not strongly block DNA synthesis or miscode in XPA cells. Primer extension from a dC terminus opposite gamma-OH-PdG was much more efficient than that opposite alpha-OH-PdG or PdG. These results indicate that the minor alpha-OH-PdG adduct is more genotoxic than the major gamma-OH-PdG. Furthermore, experiments using a HeLa whole cell extract indicate that all three DNA adducts are not efficiently removed from DNA by base excision repair.

• Authors: In-Young Yang, Grace Chan, Holly Miller, Yanhe Huang, Maria Cecilia Torres, Francis Johnson, Masaaki Moriya
• Journal: Biochemistry (Biochemistry) Vol. 41 Issue 46 Pg. 13826-32 (Nov 19 2002) ISSN: 0006-2960 [Print] United States
• PMID: 12427046 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA Adducts
• DNA Primers
• OH-1,-N(2)-propano-2'-deoxyguanosine
• 1,N(2)-propanodeoxyguanosine
• Acrolein
• Deoxyguanosine

Topics

• Acrolein (metabolism, pharmacology)
• Base Pair Mismatch
• DNA Adducts (drug effects, metabolism)
• DNA Damage
• DNA Primers (chemistry)
• DNA Repair
• DNA Replication (drug effects)
• Deoxyguanosine (analogs & derivatives, metabolism, pharmacology)
• Escherichia coli (drug effects, genetics)
• Genetic Vectors
• HeLa Cells (drug effects)
• Humans
• Mutagenesis
• Xeroderma Pigmentosum (genetics, metabolism)