A Functional 125-kDa Core Polypeptide of Fission Yeast DNA Topoisomerase II

Kazuhiro Shiozaki, Mitsuhiro Yanagida

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

We purified fission yeast DNA topoisomerase II (topo II) to apparent homogeneity. It consists of a single 165-kDa polypeptide in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and, upon treatment with a bifunctional reagent, doubles its molecular weight. Limited proteolysis of intact topo II by papain produces a 125-kDa core, which lacks the N-terminal 75 and the C-terminal ∼260 amino acids but still contains regions similar to those of bacterial or phage T4 topo II subunits. The core retains relaxing and unknotting activities. Further digestion inactivates the core, cleaving it at the middle of the GyrB-like domain and at the beginning of the GyrA-like domain. Therefore, papain appears to cleave spatially distinct subdomains of topo II. We made top2 mutant genes deleted of the C-terminal 286 or N-terminal 74 amino acids, which can substitute for the wild-type top2+ gene in mitosis and meiosis. However, a mutant containing deletions of both termini cannot rescue the top2 null mutant, despite the fact that the product is enzymatically active. Therefore, the top2 product of the doubly truncated gene may not fulfill all of the in vivo requirements for top2+ function.

Original languageEnglish (US)
Pages (from-to)6093-6102
Number of pages10
JournalMolecular and Cellular Biology
Volume11
Issue number12
StatePublished - Dec 1991
Externally publishedYes

Fingerprint

Type II DNA Topoisomerase
Schizosaccharomyces
Peptides
Papain
Cross-Linking Reagents
Genes
Amino Acids
Bacteriophage T4
Meiosis
Mitosis
Sodium Dodecyl Sulfate
Proteolysis
Polyacrylamide Gel Electrophoresis
Digestion
Molecular Weight

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

A Functional 125-kDa Core Polypeptide of Fission Yeast DNA Topoisomerase II. / Shiozaki, Kazuhiro; Yanagida, Mitsuhiro.

In: Molecular and Cellular Biology, Vol. 11, No. 12, 12.1991, p. 6093-6102.

Research output: Contribution to journalArticle

Shiozaki, Kazuhiro ; Yanagida, Mitsuhiro. / A Functional 125-kDa Core Polypeptide of Fission Yeast DNA Topoisomerase II. In: Molecular and Cellular Biology. 1991 ; Vol. 11, No. 12. pp. 6093-6102.
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