Effect of Carcinogenic Adducts on Transcription by T7 RNA Polymerase

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1987 May 26

PMID 3588293

Citations 1

Authors

S T Nath

M S Lee

L J Romano

Affiliations

Soon will be listed here.

Abstract

The effect of modifying the T7 promoter-containing plasmid pDR100 with aminofluorene (AF), acetylaminofluorene (AAF), and benzo[a]pyrene (B[a]P) adducts on RNA synthesis by the T7 RNA polymerase was determined. We found that increasing levels of each of the three adducts caused a progressive decline in RNA synthesis, but that the inhibition produced by benzo[a]pyrene adducts was substantially greater than that caused by either AF or AAF adducts. Polyacrylamide gel electrophoresis analysis confirmed that the B[a]P adducts more strongly inhibited chain elongation since their presence resulted in the production of RNA fragments having average chain lengths very close to that predicted if each adduct permanently blocked the polymerase during transcription. We have also determined the effect of each type of adduct on the initiation of transcription and show that the T7 RNA polymerase initiates as efficiently on both AF or AAF-containing templates as it does on unmodified DNA, while B[a]P adducts were found to increase in the number of chain starts.

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References

KRIEK E . On the interaction of N-2-fluorenylhydroxylamine with nucleic acids in vitro. Biochem Biophys Res Commun. 1965; 20(6):793-9. DOI: 10.1016/0006-291x(65)90088-4. View

Miller E, Juhl U, Miller J . Nucleic acid guanine: reaction with the carcinogen N-acetoxy-2-acetylaminofluorene. Science. 1966; 153(3740):1125-7. DOI: 10.1126/science.153.3740.1125. View

KRIEK E, Miller J, Juhl U, Miller E . 8-(N-2-fluorenylacetamido)guanosine, an arylamidation reaction product of guanosine and the carcinogen N-acetoxy-N-2-fluorenylacetamide in neutral solution. Biochemistry. 1967; 6(1):177-82. DOI: 10.1021/bi00853a029. View

Grunberger G, Yu F, Grunberger D, FEIGELSON P . Mechanism of N-hydroxy-2-acetylaminofluorene inhibition of rat hepatic ribonucleic acid synthesis. J Biol Chem. 1973; 248(18):6278-81. View

Levine A, Fink L, Weinstein I, Grunberger D . Effect of N-2-acetylaminofluorene modification on the conformation of nucleic acids. Cancer Res. 1974; 34(2):319-27. View

Marotta C, Lebowitz P, Dhar R, Zain B, Weissman S . Preparation of RNA transcripts of discrete segments of DNA. Methods Enzymol. 1974; 29:254-72. DOI: 10.1016/0076-6879(74)29026-8. View

MILLETTE R, Fink L . The effect of modification of T7 DNA by the carcinogen N-1-acetylaminofluorene: termination of transcription in vitro. Biochemistry. 1975; 14(7):1426-32. DOI: 10.1021/bi00678a012. View

Fuchs R . In vitro recognition of carcinogen-induced local denaturation sites native DNA by S1 endonuclease from Aspergillus oryzae. Nature. 1975; 257(5522):151-2. DOI: 10.1038/257151a0. View

Marquardt H, Grover P, Sims P . In vitro malignant transformation of mouse fibroblasts by non-K-region dihydrodiols derived from 7-methylbenz(a)anthracene, 7,12-dimethylbenz(a)anthracene, and benzo(a)pyrene. Cancer Res. 1976; 36(6):2059-64. View

10.

Newbold R, Brookes P . Exceptional mutagenicity of a benzo(a)pyrene diol epoxide in cultured mammalian cells. Nature. 1976; 261(5555):52-4. DOI: 10.1038/261052a0. View

11.

Weinstein I, Jeffrey A, Jennette K, Blobstein S, Harvey R, Harris C . Benzo(a)pyrene diol epoxides as intermediates in nucleic acid binding in vitro and in vivo. Science. 1976; 193(4253):592-5. DOI: 10.1126/science.959820. View

12.

Pulkrabek P, Leffler S, Weinstein I, Grunberger D . Conformation of DNA modified with a dihydrodiol epoxide derivative of benzo[a]pyrene. Biochemistry. 1977; 16(14):3127-32. DOI: 10.1021/bi00633a014. View

13.

Leffler S, Pulkrabek P, Grunberger D, Weinstein I . Template activity of calf thymus DNA modified by a dihydrodiol epoxide derivative of benzo[a]pyrene. Biochemistry. 1977; 16(14):3133-6. DOI: 10.1021/bi00633a015. View

14.

Jeffrey A, Weinstein I, Jennette K, Grzeskowiak K, Nakanishi K, Harvey R . Structures of benzo(a)pyrene--nucleic acid adducts formed in human and bovine bronchial explants. Nature. 1977; 269(5626):348-50. DOI: 10.1038/269348a0. View

15.

KAKEFUDA T, Yamamoto H . Modification of DNA by the benzo[a]pyrene metabolite diol-epoxide r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Proc Natl Acad Sci U S A. 1978; 75(1):415-9. PMC: 411259. DOI: 10.1073/pnas.75.1.415. View

16.

Miller E . Some current perspectives on chemical carcinogenesis in humans and experimental animals: Presidential Address. Cancer Res. 1978; 38(6):1479-96. View

17.

Drinkwater N, Miller J, Miller E, Yang N . Covalent intercalative binding to DNA in relation to the mutagenicity of hydrocarbon epoxides and N-acetoxy-2-acetylaminofluorene. Cancer Res. 1978; 38(10):3247-55. View

18.

Buening M, Wislocki P, Levin W, Yagi H, Thakker D, Akagi H . Tumorigenicity of the optical enantiomers of the diastereomeric benzo[a]pyrene 7,8-diol-9,10-epoxides in newborn mice: exceptional activity of (+)-7beta,8alpha-dihydroxy-9alpha,10alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Proc Natl Acad Sci U S A. 1978; 75(11):5358-61. PMC: 392962. DOI: 10.1073/pnas.75.11.5358. View

19.

Rinkus S, Legator M . Chemical characterization of 465 known or suspected carcinogens and their correlation with mutagenic activity in the Salmonella typhimurium system. Cancer Res. 1979; 39(9):3289-318. View

20.

Meehan T, Straub K . Double-stranded DNA steroselectively binds benzo(a)pyrene diol epoxides. Nature. 1979; 277(5695):410-2. DOI: 10.1038/277410a0. View