ATP-bound Topoisomerase II as a Target for Antitumor Drugs

Huimin Wang, Yong Mao, Nai Zhou, Tao Hu, Tao Shih Hsieh, Leroy-Fong Liu

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Topoisomerase II (TOP2) poisons interfere with the breakage/reunion reaction of TOP2 resulting in DNA cleavage. In the current studies, we show that two different classes (ATP-sensitive and -insensitive) of TOP2 poisons can be identified based on their differential sensitivity to the ATP-bound conformation of TOP2. First, in the presence of 1 mM ATP or the nonhydrolyzable analog adenosine 5′-(β-γ-imino)triphosphate, TOP2-mediated DNA cleavage induced by ATP-sensitive TOP2 poisons (e.g. doxorubicin, etoposide, mitoxantrone, and 4′-(9-acridinylamino)methanesulfon-m-anisidide) was 30-100-fold stimulated, whereas DNA cleavage induced by ATP-insensitive TOP2 poisons (e.g. amonafide, batracylin, and menadione) was only slightly (less than 3-fold) affected. In addition, ADP was shown to strongly antagonize TOP2-mediated DNA cleavage induced by ATP-sensitive but not ATP-insensitive TOP2 poisons. Second, C427A mutant human TOP2α, which exhibits reduced ATPase activity, was shown to exhibit cross-resistance to all ATP-sensitive but not ATP-insensitive TOP2 poisons. Third, using ciprofloxacin competition assay, TOP2-mediated DNA cleavage induced by ATP-sensitive but not ATP-insensitive poisons was shown to be antagonized by ciprofloxacin. These results suggest that ATP-bound TOP2 may be the specific target of ATP-sensitive TOP2 poisons. Using Lac repressor-operator complexes as roadblocks, we show that ATP-bound TOP2 acts as a circular clamp capable of entering DNA ends and sliding on unobstructed duplex DNA.

Original languageEnglish
Pages (from-to)15990-15995
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number19
DOIs
Publication statusPublished - May 11 2001
Externally publishedYes

Fingerprint

Type II DNA Topoisomerase
Antineoplastic Agents
Adenosine Triphosphate
Poisons
DNA Cleavage
DNA
batracylin
Ciprofloxacin
amonafide
Lac Repressors
Reunion
Vitamin K 3
Mitoxantrone
Clamping devices
Etoposide
Adenosine
Doxorubicin
Adenosine Diphosphate
Adenosine Triphosphatases
Conformations

ASJC Scopus subject areas

  • Biochemistry

Cite this

ATP-bound Topoisomerase II as a Target for Antitumor Drugs. / Wang, Huimin; Mao, Yong; Zhou, Nai; Hu, Tao; Hsieh, Tao Shih; Liu, Leroy-Fong.

In: Journal of Biological Chemistry, Vol. 276, No. 19, 11.05.2001, p. 15990-15995.

Research output: Contribution to journalArticle

Wang, Huimin ; Mao, Yong ; Zhou, Nai ; Hu, Tao ; Hsieh, Tao Shih ; Liu, Leroy-Fong. / ATP-bound Topoisomerase II as a Target for Antitumor Drugs. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 19. pp. 15990-15995.
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