Abstract
A key step in the Fanconi anemia (FA) tumor suppressor pathway is the site-specific monoubiquitination of the FANCD2 protein. Genetic studies indicate that this crucial modification requires eight known FA gene products and the E2-conjugating enzyme Ube2t. Here, we minimally reconstitute this monoubiquitination reaction with Ube2t and the FANCL protein, revealing that monoubiquitination is stimulated by a conserved RWD-like domain in FANCL. Furthermore, addition of the FANCI protein enhances monoubiquitination and also restricts it to the in vivo substrate lysine residue on FANCD2. This work therefore establishes a system that provides mechanistic insight into the functions of FANCL and FANCI in the catalysis of FANCD2 monoubiquitination.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Cell Line
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Chickens
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Conserved Sequence
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Fanconi Anemia Complementation Group D2 Protein / metabolism*
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Fanconi Anemia Complementation Group L Protein / chemistry
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Fanconi Anemia Complementation Group L Protein / metabolism*
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Fanconi Anemia Complementation Group Proteins / metabolism*
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Humans
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Molecular Sequence Data
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Protein Structure, Tertiary
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Substrate Specificity
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Ubiquitin-Conjugating Enzymes / metabolism*
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Ubiquitination*
Substances
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FANCI protein, human
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Fanconi Anemia Complementation Group D2 Protein
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Fanconi Anemia Complementation Group Proteins
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UBE2T protein, human
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UBE2W protein, human
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Ubiquitin-Conjugating Enzymes
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Fanconi Anemia Complementation Group L Protein