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PMID: 22902369 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

BIRC7-E2 ubiquitin conjugate structure reveals the mechanism of ubiquitin transfer by a RING dimer.

Nature structural & molecular biology ·Vol. 19 ·No. 9 ·2012-09-00 ·Pages 876-83

Dou H, Buetow L, Sibbet GJ, Cameron K, Huang DT

Abstract

Certain RING ubiquitin ligases (E3s) dimerize to facilitate ubiquitin (Ub) transfer from ubiquitin-conjugating enzyme (E2) to substrate, but structural evidence on how this process promotes Ub transfer is lacking. Here we report the structure of the human dimeric RING domain from BIRC7 in complex with the E2 UbcH5B covalently linked to Ub (UbcH5B∼Ub). The structure reveals extensive noncovalent donor Ub interactions with UbcH5B and both subunits of the RING domain dimer that stabilize the globular body and C-terminal tail of Ub. Mutations that disrupt these noncovalent interactions or RING dimerization reduce UbcH5B∼Ub binding affinity and ubiquitination activity. Moreover, NMR analyses demonstrate that BIRC7 binding to UbcH5B∼Ub induces peak-shift perturbations in the donor Ub consistent with the crystallographically-observed Ub interactions. Our results provide structural insights into how dimeric RING E3s recruit E2∼Ub and optimize the donor Ub configuration for transfer.

MeSH Terms
Adaptor Proteins, Signal Transducing/chemistry,metabolism Amino Acid Sequence Baculoviral IAP Repeat-Containing 3 Protein Crystallography, X-Ray Humans Inhibitor of Apoptosis Proteins/chemistry,metabolism Models, Molecular Molecular Sequence Data Neoplasm Proteins/chemistry,metabolism Nuclear Magnetic Resonance, Biomolecular Protein Binding Protein Conformation Protein Interaction Maps Protein Multimerization Protein Structure, Tertiary Ubiquitin/chemistry,metabolism Ubiquitin-Conjugating Enzymes/chemistry,metabolism Ubiquitin-Protein Ligases Ubiquitination
Chemicals
Adaptor Proteins, Signal Transducing BIRC7 protein, human Inhibitor of Apoptosis Proteins Neoplasm Proteins Ubiquitin UBE2D2 protein, human Ubiquitin-Conjugating Enzymes BIRC3 protein, human Baculoviral IAP Repeat-Containing 3 Protein Ubiquitin-Protein Ligases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Dou Hao
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow, G61 1BD, United Kingdom.
Buetow Lori
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow, G61 1BD, United Kingdom.
Sibbet Gary J
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow, G61 1BD, United Kingdom.
Cameron Kenneth
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow, G61 1BD, United Kingdom.
Huang Danny T
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow, G61 1BD, United Kingdom.
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Article Info
Journal
Nature structural & molecular biology
Abbr.
Nat Struct Mol Biol
ISSN
1545-9985
DOI
10.1038/nsmb.2379
Published
2012-09-00
Epub
2012-00-14
Pages
876-83
Language
English
Region
United States
NLM ID
101186374
PMCID
PMC3880866
Subset
IM
Grants
Cancer Research UK · A11593 · United Kingdom
External Links
PubMed source DOI: 10.1038/nsmb.2379 PMC Full Text View journal details
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