Cooperation of HECT-domain ubiquitin ligase hHYD and DNA topoisomerase II-binding protein for DNA damage response

Yoshiomi Honda, Masahide Tojo, Kazuhito Matsuzaki, Tadashi Anan, Mitsuhiro Matsumoto, Masayuki Ando, Hideyuki Saya, Mitsuyoshi Nakao

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

Ubiquitin ligases define the substrate specificity of protein ubiquitination and subsequent proteosomal degradation. The catalytic sequence was first characterized in the C terminus of E6-associated protein (E6AP) and referred to as the HECT (homologous to E6AP C terminus) domain. The human homologue of the regulator of cell proliferation hyperplastic discs in Drosophila, designated hHYD, is a HECT-domain ubiquitin ligase. Here we show that hHYD provides a ubiquitin system for a cellular response to DNA damage. A yeast two-hybrid screen showed that DNA topoisomerase IIβ-binding protein 1 (TopBP1) interacted with hHYD. Endogenous hHYD bound the BRCA1 C-terminus domains of TopBP1 that are highlighted in DNA damage checkpoint proteins and cell cycle regulators. Using an in vitro reconstitution, specific E2 (ubiquitin-conjugating) enzymes (human UbcH4, UbcH5B, and UbcH5C) transferred ubiquitin molecules to hHYD, leading to the ubiquitination of TopBP1. TopBP1 was usually ubiquitinated and degraded by the proteosome, whereas X-irradiation diminished the ubiquitination of TopBP1 probably via the phosphorylation, resulting in the stable colocalization of up-regulated TopBP1 with γ-H2AX nuclear foci in DNA breaks. These results demonstrated that hHYD coordinated TopBP1 in the DNA damage response.

Original languageEnglish
Pages (from-to)3599-3605
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number5
DOIs
Publication statusPublished - 2002 Feb 1
Externally publishedYes

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Type II DNA Topoisomerase
DNA-Binding Proteins
Ligases
Ubiquitin
DNA Damage
Carrier Proteins
Ubiquitination
DNA
Cell Cycle Proteins
Phosphorylation
DNA Breaks
Cell proliferation
Substrate Specificity
Cell Cycle Checkpoints
Protein C
Yeast
Drosophila
Proteins
Yeasts
Cell Proliferation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cooperation of HECT-domain ubiquitin ligase hHYD and DNA topoisomerase II-binding protein for DNA damage response. / Honda, Yoshiomi; Tojo, Masahide; Matsuzaki, Kazuhito; Anan, Tadashi; Matsumoto, Mitsuhiro; Ando, Masayuki; Saya, Hideyuki; Nakao, Mitsuyoshi.

In: Journal of Biological Chemistry, Vol. 277, No. 5, 01.02.2002, p. 3599-3605.

Research output: Contribution to journalArticle

Honda, Yoshiomi ; Tojo, Masahide ; Matsuzaki, Kazuhito ; Anan, Tadashi ; Matsumoto, Mitsuhiro ; Ando, Masayuki ; Saya, Hideyuki ; Nakao, Mitsuyoshi. / Cooperation of HECT-domain ubiquitin ligase hHYD and DNA topoisomerase II-binding protein for DNA damage response. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 5. pp. 3599-3605.
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