The Mechanism of Topoisomerase I Poisoning by a Camptothecin Analog

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2002 Nov 12

PMID 12426403

Citations 274

Authors

Bart L Staker

Kathryn Hjerrild

Michael D Feese

Craig A Behnke

Alex B Burgin Jr

Lance Stewart

Affiliations

Soon will be listed here.

Abstract

We report the x-ray crystal structure of human topoisomerase I covalently joined to double-stranded DNA and bound to the clinically approved anticancer agent Topotecan. Topotecan mimics a DNA base pair and binds at the site of DNA cleavage by intercalating between the upstream (-1) and downstream (+1) base pairs. Intercalation displaces the downstream DNA, thus preventing religation of the cleaved strand. By specifically binding to the enzyme-substrate complex, Topotecan acts as an uncompetitive inhibitor. The structure can explain several of the known structure-activity relationships of the camptothecin family of anticancer drugs and suggests that there are at least two classes of mutations that can produce a drug-resistant enzyme. The first class includes changes to residues that contribute to direct interactions with the drug, whereas a second class would alter interactions with the DNA and thereby destabilize the drug-binding site.

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References

Chang J, Liu J, Juang S, Liu T, Chen L . Novel mutation of topoisomerase I in rendering cells resistant to camptothecin. Cancer Res. 2002; 62(13):3716-21. View

Stewart L, Clark R, Behnke C . High-throughput crystallization and structure determination in drug discovery. Drug Discov Today. 2002; 7(3):187-96. DOI: 10.1016/s1359-6446(01)02121-3. View

Hertzberg R, Caranfa M, Holden K, Jakas D, Gallagher G, Mattern M . Modification of the hydroxy lactone ring of camptothecin: inhibition of mammalian topoisomerase I and biological activity. J Med Chem. 1989; 32(3):715-20. DOI: 10.1021/jm00123a038. View

Kehrer D, Soepenberg O, Loos W, Verweij J, Sparreboom A . Modulation of camptothecin analogs in the treatment of cancer: a review. Anticancer Drugs. 2001; 12(2):89-105. DOI: 10.1097/00001813-200102000-00002. View

Chourpa I, Millot J, Sockalingum G, Riou J, Manfait M . Kinetics of lactone hydrolysis in antitumor drugs of camptothecin series as studied by fluorescence spectroscopy. Biochim Biophys Acta. 1998; 1379(3):353-66. DOI: 10.1016/s0304-4165(97)00115-3. View

Fiorani P, Amatruda J, Silvestri A, Butler R, Bjornsti M, Benedetti P . Domain interactions affecting human DNA topoisomerase I catalysis and camptothecin sensitivity. Mol Pharmacol. 1999; 56(6):1105-15. DOI: 10.1124/mol.56.6.1105. View

Hsiang Y, HERTZBERG R, Hecht S, Liu L . Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem. 1985; 260(27):14873-8. View

Miller S, Pilch D . A spectrophotometric study of the pH-dependent and DNA binding properties of topotecan. Ann N Y Acad Sci. 2001; 922:309-13. DOI: 10.1111/j.1749-6632.2000.tb07051.x. View

Bailly C, Carrasco C, Hamy F, Vezin H, Prudhomme M, Saleem A . The camptothecin-resistant topoisomerase I mutant F361S is cross-resistant to antitumor rebeccamycin derivatives. A model for topoisomerase I inhibition by indolocarbazoles. Biochemistry. 1999; 38(27):8605-11. DOI: 10.1021/bi983052y. View

10.

Kerrigan J, Pilch D . A structural model for the ternary cleavable complex formed between human topoisomerase I, DNA, and camptothecin. Biochemistry. 2001; 40(33):9792-8. DOI: 10.1021/bi010913l. View

11.

Li X, Haluska Jr P, Hsiang Y, Bharti A, Kufe D, Liu L . Involvement of amino acids 361 to 364 of human topoisomerase I in camptothecin resistance and enzyme catalysis. Biochem Pharmacol. 1997; 53(7):1019-27. DOI: 10.1016/s0006-2952(96)00899-4. View

12.

Stewart L, Ireton G, Parker L, Madden K, Champoux J . Biochemical and biophysical analyses of recombinant forms of human topoisomerase I. J Biol Chem. 1996; 271(13):7593-601. DOI: 10.1074/jbc.271.13.7593. View

13.

Champoux J . DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem. 2001; 70:369-413. DOI: 10.1146/annurev.biochem.70.1.369. View

14.

Laco G, Collins J, Luke B, Kroth H, Sayer J, Jerina D . Human topoisomerase I inhibition: docking camptothecin and derivatives into a structure-based active site model. Biochemistry. 2002; 41(5):1428-35. DOI: 10.1021/bi011774a. View

15.

Sheldrick G, Schneider T . SHELXL: high-resolution refinement. Methods Enzymol. 1997; 277:319-43. View

16.

Hertzberg R, Caranfa M, Hecht S . On the mechanism of topoisomerase I inhibition by camptothecin: evidence for binding to an enzyme-DNA complex. Biochemistry. 1989; 28(11):4629-38. DOI: 10.1021/bi00437a018. View

17.

Olson W, Bansal M, Burley S, DICKERSON R, Gerstein M, Harvey S . A standard reference frame for the description of nucleic acid base-pair geometry. J Mol Biol. 2001; 313(1):229-37. DOI: 10.1006/jmbi.2001.4987. View

18.

Redinbo M, Stewart L, Champoux J, Hol W . Structural flexibility in human topoisomerase I revealed in multiple non-isomorphous crystal structures. J Mol Biol. 1999; 292(3):685-96. DOI: 10.1006/jmbi.1999.3065. View

19.

Jaxel C, Kohn K, Wani M, Wall M, Pommier Y . Structure-activity study of the actions of camptothecin derivatives on mammalian topoisomerase I: evidence for a specific receptor site and a relation to antitumor activity. Cancer Res. 1989; 49(6):1465-9. View

20.

Stewart L, Redinbo M, Qiu X, Hol W, Champoux J . A model for the mechanism of human topoisomerase I. Science. 1998; 279(5356):1534-41. DOI: 10.1126/science.279.5356.1534. View