Mechanistic insight into site-restricted monoubiquitination of FANCD2 by Ube2t, FANCL, and FANCI.

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.

• Authors: Arno F Alpi, Paul E Pace, M Madan Babu, Ketan J Patel
• Journal: Molecular cell (Mol Cell) Vol. 32 Issue 6 Pg. 767-77 (Dec 26 2008) ISSN: 1097-4164 [Electronic] United States
• PMID: 19111657 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• FANCI protein, human
• Fanconi Anemia Complementation Group D2 Protein
• Fanconi Anemia Complementation Group Proteins
• UBE2T protein, human
• UBE2W protein, human
• Ubiquitin-Conjugating Enzymes
• Fanconi Anemia Complementation Group L Protein

Topics

• Amino Acid Sequence
• Animals
• Cell Line
• Chickens
• Conserved Sequence
• Fanconi Anemia Complementation Group D2 Protein (metabolism)
• Fanconi Anemia Complementation Group L Protein (chemistry, metabolism)
• Fanconi Anemia Complementation Group Proteins (metabolism)
• Humans
• Molecular Sequence Data
• Protein Structure, Tertiary
• Substrate Specificity
• Ubiquitin-Conjugating Enzymes (metabolism)
• Ubiquitination