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

被引:152
作者
Tang, ZY
Li, B
Bharadwaj, R
Zhu, HH
Özkan, E
Hakala, K
Deisenhofer, J
Yu, HT [1 ]
机构
[1] Univ Texas, SW Med Ctr, Dept Pharmacol, Dallas, TX 75390 USA
[2] Univ Texas, SW Med Ctr, Dept Biochem, Dallas, TX 75390 USA
[3] Univ Texas, SW Med Ctr, Howard Hughes Med Inst, Dallas, TX 75390 USA
关键词
D O I
10.1091/mbc.12.12.3839
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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.
引用
收藏
页码:3839 / 3851
页数:13
相关论文
共 58 条
[1]   E2-C, a cyclin-selective ubiquitin carrier protein required for the destruction of mitotic cyclins [J].
Aristarkhov, A ;
Eytan, E ;
Moghe, A ;
Admon, A ;
Hershko, A ;
Ruderman, JV .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (09) :4294-4299
[2]   D box and KEN box motifs in budding yeast Hsl1p are required for APC-mediated degradation and direct binding to Cdc20p and Cdh1p [J].
Burton, JL ;
Solomon, MJ .
GENES & DEVELOPMENT, 2001, 15 (18) :2381-2395
[3]   SCF and cullin/RING H2-based ubiquitin ligases [J].
Deshaies, RJ .
ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 1999, 15 :435-467
[4]   Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1 [J].
Fang, GW ;
Yu, HT ;
Kirschner, MW .
MOLECULAR CELL, 1998, 2 (02) :163-171
[5]   Mdm2 is a RING finger-dependent ubiquitin protein ligase for itself and p53 [J].
Fang, SY ;
Jensen, JP ;
Ludwig, RL ;
Vousden, KH ;
Weissman, AM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (12) :8945-8951
[6]   A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p [J].
Feldman, RMR ;
Correll, CC ;
Kaplan, KB ;
Deshaies, RJ .
CELL, 1997, 91 (02) :221-230
[7]   The RING-H2 finger protein APC11 and the E2 enzyme UBC4 are sufficient to ubiquitinate substrates of the anaphase-promoting complex [J].
Gmachl, M ;
Gieffers, C ;
Podtelejnikov, AV ;
Mann, M ;
Peters, JM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (16) :8973-8978
[8]   Characterization of the DOC1/APC10 subunit of the yeast and the human anaphase-promoting complex [J].
Grossberger, R ;
Gieffers, C ;
Zachariae, W ;
Podtelejnikova, AV ;
Schleiffer, A ;
Nasmyth, K ;
Mann, M ;
Peters, JM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (20) :14500-14507
[9]   The ubiquitin system [J].
Hershko, A ;
Ciechanover, A .
ANNUAL REVIEW OF BIOCHEMISTRY, 1998, 67 :425-479
[10]   The anaphase inhibitor Pds1 binds to the APC/C-associated protein Cdc20 in a destruction box-dependent manner [J].
Hilioti, Z ;
Chung, YS ;
Mochizuki, Y ;
Hardy, CFJ ;
Cohen-Fix, O .
CURRENT BIOLOGY, 2001, 11 (17) :1347-1352