The mechanism of topoisomerase I poisoning by a camptothecin analog

November 8, 2002
99 (24) 15387-15392

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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Acknowledgments

We thank G. Ireton, E. Wallace, D. Connor, and A. Raymond for technical assistance; H. Kim and M. A. Bjornsti for intellectual support and review of this manuscript; S. Wasserman at COM-CAT (Sector 32) Advanced Photon Source, Argonne National Laboratory for assistance in data collection; and L. Berman and the staff of X25 at the National Synchrotron Light Source, Brookhaven National Laboratory, for assistance in data collection and helpful support. This work was supported by National Cancer Institute Grants R43 CA82964 and R43 CA79439; by the Pacific West Cancer Fund; and by National Institute for General Medicine Grant GM58598.

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 99 | No. 24
November 26, 2002
PubMed: 12426403

Classifications

Submission history

Received: May 1, 2002
Published online: November 8, 2002
Published in issue: November 26, 2002

Acknowledgments

We thank G. Ireton, E. Wallace, D. Connor, and A. Raymond for technical assistance; H. Kim and M. A. Bjornsti for intellectual support and review of this manuscript; S. Wasserman at COM-CAT (Sector 32) Advanced Photon Source, Argonne National Laboratory for assistance in data collection; and L. Berman and the staff of X25 at the National Synchrotron Light Source, Brookhaven National Laboratory, for assistance in data collection and helpful support. This work was supported by National Cancer Institute Grants R43 CA82964 and R43 CA79439; by the Pacific West Cancer Fund; and by National Institute for General Medicine Grant GM58598.

Authors

Affiliations

Bart L. Staker
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110
Kathryn Hjerrild
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110
Michael D. Feese
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110
Craig A. Behnke*
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110
Alex B. Burgin, Jr.
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110
Lance Stewart
deCODE genetics, Incorporated, BioStructures Group, 7869 Northeast Day Road West, Bainbridge Island, WA 98110

Notes

*
Present address: Syrrx, Inc., 10450 Science Center Drive, Suite 100, San Diego, CA 92121.
To whom correspondence should be addressed. E-mail: [email protected].
Edited by James C. Wang, Harvard University, Cambridge, MA, and approved October 2, 2002

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    The mechanism of topoisomerase I poisoning by a camptothecin analog
    Proceedings of the National Academy of Sciences
    • Vol. 99
    • No. 24
    • pp. 15247-15831

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