Structural analysis of human FANCL, the E3 ligase in the Fanconi anemia pathway.
Structural analysis of human FANCL, the E3 ligase in the Fanconi anemia pathway.
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DOI：
10.1074/jbc.m111.244632
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发表时间：
2011-09-16
期刊：
影响因子：
--
通讯作者：
Walden H
中科院分区：
文献类型：
--
作者：
Hodson C;Cole AR;Lewis LP;Miles JA;Purkiss A;Walden H
范可尼贫血（FA）通路对于DNA链间交联的修复是必不可少的。该途径的核心是通过含有E3泛素连接酶FANCL的多蛋白“核心复合物”对FANCI-FANCD 2（ID）复合物的单泛素化。脊椎动物有八个蛋白质的核心复合物，而无脊椎动物显然没有。我们在这里报告的人类FANCL的中央结构域的结构与最近解决的果蝇FANCL。我们的数据代表了人类系统催化核心的第一个结构细节，并揭示了FANCL的中心折叠在物种之间是保守的。然而，FANCL蛋白之间存在大分子差异，这可能是脊椎动物和无脊椎动物FA途径之间核心复合物需求的明显区别。此外，我们表征了人FANCL与其伙伴Ube 2 t、FANCD 2和FANCI的结合。突变分析揭示了底物结合所需的残基，我们还显示了E2结合所需的结构域。
The Fanconi anemia (FA) pathway is essential for the repair of DNA interstrand cross-links. At the heart of this pathway is the monoubiquitination of the FANCI-FANCD2 (ID) complex by the multiprotein “core complex” containing the E3 ubiquitin ligase FANCL. Vertebrate organisms have the eight-protein core complex, whereas invertebrates apparently do not. We report here the structure of the central domain of human FANCL in comparison with the recently solved Drosophila melanogaster FANCL. Our data represent the first structural detail into the catalytic core of the human system and reveal that the central fold of FANCL is conserved between species. However, there are macromolecular differences between the FANCL proteins that may account for the apparent distinctions in core complex requirements between the vertebrate and invertebrate FA pathways. In addition, we characterize the binding of human FANCL with its partners, Ube2t, FANCD2, and FANCI. Mutational analysis reveals which residues are required for substrate binding, and we also show the domain required for E2 binding.