Base sequence effects in bending induced by bulky carcinogen - DNA adducts

Experimental and computational analysis

Q. Ruan, P. Zhuang, S. Li, R. Perlow, A. R. Srinivasan, X. J. Lu, S. Broyde, W. K. Olson, N. E. Geacintov

Research output: Contribution to journalArticle

Abstract

The covalent binding of bulky mutagenic or carcinogenic compounds to DNA can lead to bending, which could significantly alter the interactions of DNA with critical replication and transcription proteins. The impact of adducts derived from the highly reactive bay region enantiomeric (+)- and (-)-anti-7,8-diol-9,10-epoxide derivatives of benzo[a]pyrene (BPDE) are of interest because the (+)-7R,8S,9S,10R-anti-BPDE enantiomer is highly tumorigenic in rodents, while the (-)-7S,8R,9R, 10S-anti-BPDE enantiomer is not. Both (+)- and (-)-anti-BPDE bind covalently with DNA predominantly by trans addition at the exocyclic amino group of guanine to yield 10S (+)- and 10R (-)-trans-anti-[BP]-N 2-dG adducts. We have synthesized a number of different oligonucleotides with single (+)- and (-)-trans-anti-[BP]-N 2-dG adducts (G*) in the base sequence context XG*Y, where X and Y are different DNA bases. The G* residues were positioned at or close to the center of 11 base pair (∼1 helical turn) or 16 base pair (∼1.5 turns) duplexes. All bases, except for X and Y and their partners, were identical. These sequences were self-ligated with T4 ligase to form multimers that yield a ladder of bands upon electrophoresis in native polyacrylamide gels. The extent of bending in each oligonucleotide was assessed by monitoring the decrease in gel mobilities of these linear, self-ligated oligomers, relative to unmodified oligonucleotides of the same base sequence. The extent of global bending was then estimated using a sequence-specific three-dimensional model from which the values of the base-pair step parameter roll adjacent to the lesion site could be extracted. We find that (+)-trans-anti-[BP]-N 2-dG adducts are considerably more bent than the (-) isomers regardless of sequence and that A-T base pairs flanking the [BP]-N 2-dG lesion site allow for local flexibility consistent with adduct conformational heterogeneity. Interestingly, the fit of computed versus observed gel mobilities using classical reptation treatments requires enhancement of unmodified DNA flexibility in gels, compared to aqueous salt solution. The differences in bending between the two stereoisomeric adduct duplexes and the observed base sequence context effects may play a significant role in the differential processing of these lesions by cellular replication, transcription, and repair enzymes.

Original languageEnglish (US)
Pages (from-to)10458-10472
Number of pages15
JournalBiochemistry
Volume40
Issue number35
DOIs
StatePublished - Sep 4 2001

Fingerprint

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
DNA Adducts
Base Pairing
Carcinogens
Oligonucleotides
DNA
Enantiomers
Gels
Transcription
Benzo(a)pyrene
Epoxy Compounds
Ladders
Guanine
Ligases
Electrophoresis
Oligomers
Isomers
Native Polyacrylamide Gel Electrophoresis
Repair
Salts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Base sequence effects in bending induced by bulky carcinogen - DNA adducts : Experimental and computational analysis. / Ruan, Q.; Zhuang, P.; Li, S.; Perlow, R.; Srinivasan, A. R.; Lu, X. J.; Broyde, S.; Olson, W. K.; Geacintov, N. E.

In: Biochemistry, Vol. 40, No. 35, 04.09.2001, p. 10458-10472.

Research output: Contribution to journalArticle

Ruan, Q. ; Zhuang, P. ; Li, S. ; Perlow, R. ; Srinivasan, A. R. ; Lu, X. J. ; Broyde, S. ; Olson, W. K. ; Geacintov, N. E. / Base sequence effects in bending induced by bulky carcinogen - DNA adducts : Experimental and computational analysis. In: Biochemistry. 2001 ; Vol. 40, No. 35. pp. 10458-10472.
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T2 - Experimental and computational analysis

AU - Ruan, Q.

AU - Zhuang, P.

AU - Li, S.

AU - Perlow, R.

AU - Srinivasan, A. R.

AU - Lu, X. J.

AU - Broyde, S.

AU - Olson, W. K.

AU - Geacintov, N. E.

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