Dcn1 functions as a scaffold-type E3 ligase for cullin neddylation
Dcn1 functions as a scaffold-type E3 ligase for cullin neddylation
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DOI：
10.1016/j.molcel.2007.12.012
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发表时间：
2008-01-18
期刊：
影响因子：
16
通讯作者：
Sicheri, Frank
中科院分区：
文献类型：
--
作者：
Kurz, Thimo;Chou, Yang-Chieh;Sicheri, Frank
基于cullin的E3泛素连接酶通过用泛素样蛋白Nedd 8修饰cullin亚基而被激活。dcn 1调节cullin neddylation和泛素连接酶活性。在这里，我们描述了酵母Dcn 1的1.9埃的X射线晶体结构，包括一个N-末端泛素结合（乌巴）结构域和一个C-末端结构域的独特架构，我们称之为PONY结构域。Dcn 1上的保守表面需要直接结合到cullin和neddylation。Cdc 53上Dcn 1的相互结合位点位于离neddylation位点18埃的位置。Dcn 1不需要半胱氨酸残基的催化功能，并直接与Nedd 8 E2 Ubc 12的E1结合位点重叠的表面上相互作用。我们表明，DCN 1是必要的和足够的cullin neddylation在一个纯化的重组系统。综上所述，这些数据表明，Dcn 1是一个支架样的E3连接酶的cullin neddylation。
Cullin-based E3 ubiquitin ligases are activated through modification of the cullin subunit with the ubiquitin-like protein Nedd8. Dcn1 regulates cullin neddylation and thus ubiquitin ligase activity. Here we describe the 1.9 angstrom X-ray crystal structure of yeast Dcn1 encompassing an N-terminal ubiquitin-binding (UBA) domain and a C-terminal domain of unique architecture, which we termed PONY domain. A conserved surface on Dcn1 is required for direct binding to cullins and for neddylation. The reciprocal binding site for Dcn1 on Cdc53 is located similar to 18 angstrom from the site of neddylation. Dcn1 does not require cysteine residues for catalytic function, and directly interacts with the Nedd8 E2 Ubc12 on a surface that overlaps with the E1-binding site. We show that Dcn1 is necessary and sufficient for cullin neddylation in a purified recombinant system. Taken together, these data demonstrate that Dcn1 is a scaffold-like E3 ligase for cullin neddylation.