Journal article
Mechanism of ubiquitin ligation and lysine prioritization by a HECT E3
eLife, Vol.2, pp.e00828-e00828
08/08/2013
DOI: 10.7554/eLife.00828
PMCID: PMC3738095
PMID: 23936628
Abstract
Ubiquitination by HECT E3 enzymes regulates myriad processes, including tumor suppression, transcription, protein trafficking, and degradation. HECT E3s use a two-step mechanism to ligate ubiquitin to target proteins. The first step is guided by interactions between the catalytic HECT domain and the E2∼ubiquitin intermediate, which promote formation of a transient, thioester-bonded HECT∼ubiquitin intermediate. Here we report that the second step of ligation is mediated by a distinct catalytic architecture established by both the HECT E3 and its covalently linked ubiquitin. The structure of a chemically trapped proxy for an E3∼ubiquitin-substrate intermediate reveals three-way interactions between ubiquitin and the bilobal HECT domain orienting the E3∼ubiquitin thioester bond for ligation, and restricting the location of the substrate-binding domain to prioritize target lysines for ubiquitination. The data allow visualization of an E2-to-E3-to-substrate ubiquitin transfer cascade, and show how HECT-specific ubiquitin interactions driving multiple reactions are repurposed by a major E3 conformational change to promote ligation. DOI:http://dx.doi.org/10.7554/eLife.00828.001.
Details
- Title: Subtitle
- Mechanism of ubiquitin ligation and lysine prioritization by a HECT E3
- Creators
- Hari B Kamadurai - St. Jude Children's Research HospitalYu Qiu - St. Jude Children's Research HospitalAlan Deng - St. Jude Children's Research HospitalJoseph S Harrison - University of North Carolina at Chapel HillChris Macdonald - University of IowaMarcelo Actis - St. Jude Children's Research HospitalPatrick Rodrigues - St. Jude Children's Research HospitalDarcie J Miller - St. Jude Children's Research HospitalJudith Souphron - Institute CurieSteven M Lewis - University of North Carolina at Chapel HillIgor Kurinov - Cornell UniversityNaoaki Fujii - St. Jude Children's Research HospitalMichal Hammel - Lawrence Berkeley National LaboratoryRobert Piper - University of IowaBrian Kuhlman - University of North Carolina at Chapel HillBrenda A Schulman - St. Jude Children's Research Hospital
- Resource Type
- Journal article
- Publication Details
- eLife, Vol.2, pp.e00828-e00828
- DOI
- 10.7554/eLife.00828
- PMID
- 23936628
- PMCID
- PMC3738095
- NLM abbreviation
- Elife
- ISSN
- 2050-084X
- eISSN
- 2050-084X
- Grant note
- 8P41 GM103403-10 / NIGMS NIH HHS 5P41RR015301-10 / NCRR NIH HHS 5P30CA021765 / NCI NIH HHS R01 GM069530 / NIGMS NIH HHS 5R01 GM058202 / NIGMS NIH HHS Howard Hughes Medical Institute R01 GM073960 / NIGMS NIH HHS R01 GM058202 / NIGMS NIH HHS P30 CA021765 / NCI NIH HHS T32 CA009156 / NCI NIH HHS R01GM069530 / NIGMS NIH HHS R01GM073960 / NIGMS NIH HHS
- Language
- English
- Date published
- 08/08/2013
- Academic Unit
- Molecular Physiology and Biophysics; Medicine Administration; Internal Medicine
- Record Identifier
- 9984297502602771
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