Oxidative thymine dimer repair in the DNA helix

Science

Volumn 275, Issue 5305, 1997, Pages 1465-1468

  Dandliker, Peter J ^a   Erik Holmlin, R ^a   Barton, Jacqueline K ^a  

^a California Institute of Technology ^*  (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DNA;

ARTICLE; DNA DAMAGE; DNA HELIX; DNA REPAIR; PHOTOCHEMISTRY; PRIORITY JOURNAL; ULTRAVIOLET RADIATION;

2,2'-DIPYRIDYL; BASE COMPOSITION; DNA; DNA DAMAGE; DNA REPAIR; ELECTRON TRANSPORT; ELECTRONS; INTERCALATING AGENTS; LIGHT; NUCLEIC ACID CONFORMATION; ORGANOMETALLIC COMPOUNDS; OXIDATION-REDUCTION; PYRIMIDINE DIMERS; RHODIUM;

EID: 1842330888     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5305.1465     Document Type: Article

References (43)

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19. 3+./2+), of ∼2.0 V versus normal hydrogen electrode in dimethylformamide (DMF), which should be sufficient to oxidize the thymine dimer [C. Turro, A. Evenzahav, S. H. Bossmann, J. K. Barton, N. J. Turro, Inorg. Chim. Acta 243, 101 (1996)].
20. Thymine dimer-containing oligonucleotides are prepared by photodimerization of single-stranded DNA at 330 nm with acetophenone as a sensitizer. The chromatogram of the photoproducts and the product ratios are consistent with those reported earlier and support the assignment of the major product as the cis/syn isomer [S. K. Banerjee, R. B. Christensen, C. W. Lawrence, J. E. LeClerc, Proc. Natl. Acad. Sci. U.S.A. 85, 8141 (1988)]. The products were characterized by enzymatic digestion from the 3′ end by T4-DNA polymerase (which showed that cleavage did not proceed past the site of dimer incorporation) and by direct photocycloreversion of the dimer-containing strands with 254-nm light to regenerate the undamaged strand.
21. 3+ to the DNA duplex.
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28. After 6 hours of irradiation at 400 nm, 5% repair of the thymine dimer was found in control samples lacking rhodium (light control) and represents the background repair by light in our experiments.
29. -1.
30. Photocleavage reactions at 313 nm, measured by phosphoimagery, sensitively detect strand cleavage at several orders of magnitude lower intensity than required to detect rhodium reaction near the tethered end of the duplex in Rh-modified oligomers. Photocleavage reactions as a function of concentration of Rh-modified duplexes also indicate that interduplex reaction is negligible at duplex concentrations of ≤25 μM.
31. Assuming a 3.4 Å centroid-to-centroid distance between stacked base pairs, the separation between the intercalated phi ligand of the rhodium complex and the center of the thymine dimer is 19 Å for both the 5′-Rh-and 3′-Rh-tethered assemblies when the complex is intercalated between the third and fourth base pair from the end of the duplex. On the basis of the photocleavage pattern (Fig. 2), the closest distance between intercalator and thymine dimer along the helical axis appears to be 16 Å for both assemblies with intercalation in the fourth base step.
32. For both diastereomers, direct photocleavage experiments (313 nm) show the metal complex to be fully bound by intercalation at these concentrations.
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35. Nuclear magnetic resonance studies have suggested somewhat greater perturbations of the thymine dimer to its 3′ side [J. Kemmink et al., Nucleic Acids Res. 15, 4645 (1987); J.-S. Taylor, D. S. Garrett, I. R. Brockie, D. L. Svoboda, J. Telser, Biochemistry 29, 8858 (1990)].
36. 2+ intercalated into the unmodified duplex (without thymine dimer). For the corresponding 5′-tethered rhodium duplex, luminescence quenching was 58% and 52% for Δ-Rh and Λ-Rh, respectively. The higher repair yields therefore correlate with higher quenching of ruthenium luminescence.
37. An analogous sensitivity in long-range oxidation of guanine doublets to intervening bulges in DNA has been observed in our laboratory (D. Hall and J. K. Barton, unpublished results).
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40. 3+ on the duplex containing a thymine dimer do not suggest preferential binding of the rhodium adjacent to the dimer.
41. 3+ and by 5°C with Δ-Rh tethered to the 5′-terminus.
42. D. G. Vassylyev et al., Cell 83, 773 (1995).
43. Supported by NIH grant GM49216, a postdoctoral fellowship from the Cancer Research Fund of the Damon Runyon-Walter Winchell Foundation (P.J.D.), and an NSF predoctoral fellowship (R.E.H.). We thank N. J. Turro and E. D. A. Stemp for helpful discussions.