Functional dissection of the phosphorylated termini of fission yeast DNA topoisomerase II

Kazuhiro Shiozaki, Mitsuhiro Yanagida

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76 Citations (Scopus)

Abstract

Fission Yeast DNA topoisomerase II (165 kD) consists of an enzymatically active 125-kD core, ∼10-kD NH2-terminal and 30-kD COOH-terminal domains. The question addressed in the present study is what is the role of the topo II termini. Although deletion of either the NH2 or the COOH terminus is viable, deletion of both termini is lethal; the termini share an essential role for viability. We show here that topo II phosphorylation sites are localized in the terminal domains, but dephosphorylated topo II is still active. The topo II terminal sequences are required for nuclear localization; topo II double terminal deletion mutants are deficient for nuclear targeting, whereas wild-type and single deletion mutant topo IIs are transported into the nucleus with different efficiencies. Functional subdomains in the NH2 terminus are further dissected; we identified a 15 amino acid nuclear localization sequence (NLS) which is essential for viability and nuclear localization when the COOH terminus is deleted. This NLS could be substituted with SV-40 large T-antigen NLS. Two other functional subdomains were found; a non-essential acidic stretch which is phosphorylated and apparently enhances the nuclear localization and an essential hydrophilic stretch of unknown function. Motifs similar to these three NH2-terminal subdomains are also found in the COOH terminus. Our results support the possibility that phosphorylation of topo II does not play an essential role in fission yeast.

Original languageEnglish (US)
Pages (from-to)1023-1036
Number of pages14
JournalJournal of Cell Biology
Volume119
Issue number5
StatePublished - Dec 1992
Externally publishedYes

Fingerprint

Type II DNA Topoisomerase
Schizosaccharomyces
Dissection
Phosphorylation
Viral Tumor Antigens
Amino Acids

ASJC Scopus subject areas

  • Cell Biology

Cite this

Functional dissection of the phosphorylated termini of fission yeast DNA topoisomerase II. / Shiozaki, Kazuhiro; Yanagida, Mitsuhiro.

In: Journal of Cell Biology, Vol. 119, No. 5, 12.1992, p. 1023-1036.

Research output: Contribution to journalArticle

Shiozaki, Kazuhiro ; Yanagida, Mitsuhiro. / Functional dissection of the phosphorylated termini of fission yeast DNA topoisomerase II. In: Journal of Cell Biology. 1992 ; Vol. 119, No. 5. pp. 1023-1036.
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