Binding of ATP to human DNA topoisomerase I resulting in an alteration of the conformation of the enzyme

Hui Jye Chen, Jaulang Hwang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

It has been known for some time that ATP inhibits the DNA relaxation activity of human DNA topoisomerase I. However, the underlying mechanism of this inhibitory effect remains largely unknown. Using filter binding assays, the binding of human DNA topoisomerase I to DNA was decreased in the presence of ATP. This result suggests that the inhibition of DNA relaxation activity of human DNA topoisomerase I by ATP is at the binding step rather than at the nicking or resealing step. DNA topoisomerase I cleavage assay further supports this notion. ATP-agarose binding and UV cross-linking assays also demonstrate that ATP directly and specifically binds human DNA topoisomerase I. To address whether the ATP binding results in conformational changes in human DNA topoisomerase I, various proteases were employed for detecting potential protein conformational changes. Our results indicated that the proteolytic susceptibilities of trypsin and chymotrypsin were altered in the presence of ATP. The result suggests that the conformation of human DNA topoisomerase I was altered upon ATP binding. In addition, the binding between ATP and human DNA topoisomerase I was also reduced by increasing concentrations of DNA. Our data suggests that human DNA topoisomerase I exhibits at least two incompatible conformations. One conformation is in the form of a topoisomerase I-ATP complex, which inhibits DNA relaxation activity of human DNA topoisomerase I, and the other, a topoisomerase I-DNA complex, which exerts DNA relaxation activity. Our studies identify the role of ATP in the regulation of human DNA topoisomerase I and provide a substantial implication of how human DNA topoisomerase I compromises its versatile functions.

Original languageEnglish
Pages (from-to)367-375
Number of pages9
JournalEuropean Journal of Biochemistry
Volume265
Issue number1
DOIs
Publication statusPublished - Oct 1 1999
Externally publishedYes

Fingerprint

Conformations
Adenosine Triphosphate
Enzymes
Type I DNA Topoisomerase
DNA
Assays
human TOP1 protein
Peptide Hydrolases

Keywords

  • ATP
  • ATP-inhibitory effect
  • Conformational change
  • DNA topoisomerase I
  • Dual functions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding of ATP to human DNA topoisomerase I resulting in an alteration of the conformation of the enzyme. / Chen, Hui Jye; Hwang, Jaulang.

In: European Journal of Biochemistry, Vol. 265, No. 1, 01.10.1999, p. 367-375.

Research output: Contribution to journalArticle

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