APC2 cullin protein and APC11 RING protein comprise the minimal ubiquitin ligase module of the anaphase-promoting complex

Z. Tang, B. Li, R. Bharadwaj, H. Zhu, E. Özkan, K. Hakala, J. Deisenhofer, H. Yu

Research output: Contribution to journalArticle

134 Citations (Scopus)

Abstract

In mitosis, the anaphase-promoting complex (APC) regulates the onset of sister-chromatid separation and exit from mitosis by mediating the ubiquitination and degradation of the securin protein and mitotic cyclins. With the use of a baculoviral expression system, we have reconstituted the ubiquitin ligase activity of human APC. In combination with Ubc4 or UbcH10, a heterodimeric complex of APC2 and APC11 is sufficient to catalyze the ubiquitination of human securin and cyclin B1. However, the minimal APC2/11 ubiquitin ligase module does not possess substrate specificity, because it also ubiquitinates the destruction box deletion mutants of securin and cyclin B1. Both APC11 and UbcH10 bind to the C-terminal cullin homology domain of APC2, whereas Ubc4 interacts with APC11 directly. Zn2+-binding and mutagenesis experiments indicate that APC11 binds Zn2+ at a 1:3 M ratio. Unlike the two Zn2+ ions of the canonical RING-finger motif, the third Zn2+ ion of APC11 is not essential for its ligase activity. Surprisingly, with Ubc4 as the E2 enzyme, Zn2+ ions alone are sufficient to catalyze the ubiquitination of cyclin B1. Therefore, the Zn2+ ions of the RING finger family of ubiquitin ligases may be directly involved in catalysis.

Original languageEnglish (US)
Pages (from-to)3839-3851
Number of pages13
JournalMolecular Biology of the Cell
Volume12
Issue number12
StatePublished - 2001

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Cullin Proteins
Anaphase-Promoting Complex-Cyclosome
Ligases
Ubiquitin
Cyclin B1
Ubiquitination
Securin
Ions
Mitosis
Proteins
RING Finger Domains
Chromatids
Cyclins
Substrate Specificity
Catalysis
Human Activities
Mutagenesis
Proteolysis
Enzymes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

APC2 cullin protein and APC11 RING protein comprise the minimal ubiquitin ligase module of the anaphase-promoting complex. / Tang, Z.; Li, B.; Bharadwaj, R.; Zhu, H.; Özkan, E.; Hakala, K.; Deisenhofer, J.; Yu, H.

In: Molecular Biology of the Cell, Vol. 12, No. 12, 2001, p. 3839-3851.

Research output: Contribution to journalArticle

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AU - Tang, Z.

AU - Li, B.

AU - Bharadwaj, R.

AU - Zhu, H.

AU - Özkan, E.

AU - Hakala, K.

AU - Deisenhofer, J.

AU - Yu, H.

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