DNA-binding activity of rat DNA topoisomerase II α C-terminal domain contributes to efficient DNA catenation in vitro

Shinji Kawano, Yuri Kato, Natsumi Okada, Kuniaki Sano, Ken Tsutsui, Kimiko M. Tsutsui, Shogo Ikeda

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

DNA topoisomerase IIα (topo IIα) is an essential enzyme for resolution of DNA topologies arising in DNA metabolic reactions. In proliferating cells, topo II activities of DNA catenation or decatenation are required for condensation of chromosomes and segregation of chromatids. Recent studies suggest that the C-terminal domain (CTD) of human topo IIα is required for localization to mitotic chromosomes. Here, we show that the CTD of topo IIα is also associated with efficient DNA catenation in vitro, based on comparison of wild-type (WT) rat topo IIα and its deletion mutants. Unlike WT, the CTD truncated mutant (ΔCTD) lacked linear DNA binding activity, but could bind to negatively supercoiled DNA similarly to WT. The CTD alone showed linear DNA-binding activity. ΔCTD mediated formation of a DNA catenane in the presence of polyethylene glycol, which enhances macromolecular association. These results indicate that DNA-binding activity in the CTD of topo IIα concentrates the enzyme in the vicinity of condensed DNA and allows topo IIα to efficiently form a DNA catenane.

Original languageEnglish
Pages (from-to)363-369
Number of pages7
JournalJournal of Biochemistry
Volume159
Issue number3
DOIs
Publication statusPublished - Aug 24 2015

Fingerprint

Type II DNA Topoisomerase
Rats
DNA
Catenated DNA
Chromosomes
Superhelical DNA
Chromosome Segregation
Chromatids
Enzymes
In Vitro Techniques
Condensation
Topology
Association reactions

Keywords

  • C-terminal domain
  • catenation
  • DNA-binding
  • topoisomerase II
  • α isoform

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

DNA-binding activity of rat DNA topoisomerase II α C-terminal domain contributes to efficient DNA catenation in vitro. / Kawano, Shinji; Kato, Yuri; Okada, Natsumi; Sano, Kuniaki; Tsutsui, Ken; Tsutsui, Kimiko M.; Ikeda, Shogo.

In: Journal of Biochemistry, Vol. 159, No. 3, 24.08.2015, p. 363-369.

Research output: Contribution to journalArticle

Kawano, Shinji ; Kato, Yuri ; Okada, Natsumi ; Sano, Kuniaki ; Tsutsui, Ken ; Tsutsui, Kimiko M. ; Ikeda, Shogo. / DNA-binding activity of rat DNA topoisomerase II α C-terminal domain contributes to efficient DNA catenation in vitro. In: Journal of Biochemistry. 2015 ; Vol. 159, No. 3. pp. 363-369.
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