Molecular determinants of polyubiquitin linkage selection by an HECT ubiquitin ligase
Open Access
- 6 April 2006
- journal article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 25 (8), 1710-1719
- https://doi.org/10.1038/sj.emboj.7601061
Abstract
Ubiquitin (Ub)‐protein ligases (E3s) frequently modify their substrates with multiple Ub molecules in the form of a polyubiquitin (poly‐Ub) chain. Although structurally distinct poly‐Ub chains (linked through different Ub lysine (Lys) residues) can confer different fates on target proteins, little is known about how E3s select the Lys residue to be used in chain synthesis. Here, we used a combination of mutagenesis, biochemistry, and mass spectrometry to map determinants of linkage choice in chain assembly catalyzed by KIAA10, an HECT (Homologous to E6AP C‐Terminus) domain E3 that synthesizes K29‐ and K48‐linked chains. Focusing on the Ub molecule that contributes the Lys residue for chain formation, we found that specific surface residues adjacent to K48 and K29 are critical for the usage of the respective Lys residues in chain synthesis. This direct mechanism of linkage choice bears similarities to the mechanism of substrate site selection in sumoylation catalyzed by Ubc9, but is distinct from the mechanism of chain linkage selection used by the Mms2/Ubc13 (Ub E2 variant (UEV)/E2) complex.Keywords
This publication has 50 references indexed in Scilit:
- Different HECT domain ubiquitin ligases employ distinct mechanisms of polyubiquitin chain synthesisThe EMBO Journal, 2005
- Insights into E3 ligase activity revealed by a SUMO–RanGAP1–Ubc9–Nup358 complexNature, 2005
- Function and regulation of cullin–RING ubiquitin ligasesNature Reviews Molecular Cell Biology, 2005
- A proteomics approach to understanding protein ubiquitinationNature Biotechnology, 2003
- Structural Basis for E2-Mediated SUMO Conjugation Revealed by a Complex between Ubiquitin-Conjugating Enzyme Ubc9 and RanGAP1Cell, 2002
- Distinct Functional Surface Regions on UbiquitinJournal of Biological Chemistry, 2001
- Ubiquitin Lys63 is involved in ubiquitination of a yeast plasma membrane proteinThe EMBO Journal, 1997
- A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase.Proceedings of the National Academy of Sciences of the United States of America, 1995
- Protein ubiquitination involving an E1–E2–E3 enzyme ubiquitin thioester cascadeNature, 1995
- An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein databaseJournal of the American Society for Mass Spectrometry, 1994