The α4 residues of human DNA topoisomerase IIα function in enzymatic activity and anticancer drug sensitivity

Namiko Suda, Yasutomo Ito, Tsuneo Imai, Toyone Kikumori, Akihiko Kikuchi, Yukihiro Nishiyama, Shonen Yoshida, Motoshi Suzuki

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We introduced a series of Pro substitutions within and near the α4 helix, a part of the breakage/rejoining region, in human DNA topoisomerase IIα, and analyzed if this region is involved in determination of anti-cancer drug sensitivity in a temperature-sensitive yeast strain (top2-4 allele). Among the 19 mutants generated, H759P and N770P showed resistance to etoposide and doxorubicin at the non-permissive temperature, where cell growth depends on activity of the human enzyme. For these residues, mutants with an Ala substitution were further created, in which H759A also showed resistance to etoposide. H759P, H759A and N770P were expressed, purified and subjected to in vitro measurement of drug sensitivity. They generated lower amounts of the etoposide-induced cleavable complexes, and were also found to have lower decatenation activity than the wild-type. In the crystal structure, the yeast equivalent of His759 is found in the vicinity of the Arg713, a putative anchoring residue of the 3′-side of cleaved DNA strands. These results suggest that His759 and the other α4 helix residues are involved in the enzymatic activity and drug sensitivity of human DNA topoisomerase IIα, via interaction with cleaved DNA.

Original languageEnglish
Pages (from-to)1767-1773
Number of pages7
JournalNucleic Acids Research
Volume32
Issue number5
DOIs
Publication statusPublished - 09-07-2004
Externally publishedYes

Fingerprint

Type II DNA Topoisomerase
Etoposide
Yeasts
Pharmaceutical Preparations
Temperature
DNA
Human Activities
Doxorubicin
Alleles
Enzymes
Growth
Neoplasms

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Suda, Namiko ; Ito, Yasutomo ; Imai, Tsuneo ; Kikumori, Toyone ; Kikuchi, Akihiko ; Nishiyama, Yukihiro ; Yoshida, Shonen ; Suzuki, Motoshi. / The α4 residues of human DNA topoisomerase IIα function in enzymatic activity and anticancer drug sensitivity. In: Nucleic Acids Research. 2004 ; Vol. 32, No. 5. pp. 1767-1773.
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The α4 residues of human DNA topoisomerase IIα function in enzymatic activity and anticancer drug sensitivity. / Suda, Namiko; Ito, Yasutomo; Imai, Tsuneo; Kikumori, Toyone; Kikuchi, Akihiko; Nishiyama, Yukihiro; Yoshida, Shonen; Suzuki, Motoshi.

In: Nucleic Acids Research, Vol. 32, No. 5, 09.07.2004, p. 1767-1773.

Research output: Contribution to journalArticle

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AU - Ito, Yasutomo

AU - Imai, Tsuneo

AU - Kikumori, Toyone

AU - Kikuchi, Akihiko

AU - Nishiyama, Yukihiro

AU - Yoshida, Shonen

AU - Suzuki, Motoshi

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