Binding and recognition in the assembly of an active BRCA1/BARD1 ubiquitin-ligase complex
Open accessAbstract
BRCA1 is a breast and ovarian cancer tumor suppressor protein that associates with BARD1 to form a RINGRING heterodimer. The BRCA1BARD1 RING complex functions as an ubiquitin (Ub) ligase with activity substantially greater than individual BRCA1 or BARD1 subunits. By using NMR spectroscopy and site-directed mutagenesis, we have mapped the binding site on the BRCA1BARD1 heterodimer for the Ub-conjugating enzyme UbcH5c. The results demonstrate that UbcH5c binds only to the BRCA1 RING domain and not the BARD1 RING. The binding interface is formed by the first and second Zn(2+)-loops and central alpha-helix of the BRCA1 RING domain, a region disrupted by cancer-predisposing mutations. Unexpectedly, a second Ub-conjugating enzyme, UbcH7, also interacts with the BRCA1BARD1 complex with similar affinity, although it is not active in Ub-ligase activity assays. Thus, binding alone is not sufficient for BRCA1-dependent Ub-ligase activity.
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In the Brca1-Bard1 heterodimer, E2 binds only to the Brca1 RING, at residues proximal to but not within the dimerization interface.
“In the context of the Brca1–Bard1 heterodimer, E2 binds only to the Brca1 RING, at residues proximal to (but not within) the dimerization interface [19].”
From The p53 orchestra: Mdm2 and Mdmx set the tone (2010) · cites this paper as [19]
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- Authors
- Brzovic, P. S., Keeffe, J. R., Nishikawa, H., Miyamoto, K., Fox, D., Fukuda, M., Ohta, T., & Klevit, R.
- Affiliations
- Department of Biochemistry, University of Washington, Seattle, WA 98195; and Division of Breast and Endocrine Surgery, St. Marianna University School of Medicine, Kawasaki 216-8511, Japan
- St. Marianna University School of Medicine (JP)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2003-05-05
- Volume
- 100
- Issue
- 10
- Pages
- 5646-5651
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC156255
- OpenAlex
- W1969807455
- Cited by
- 346
- References
- 41
- Citations by year
- 2026: 5 · 2025: 8 · 2024: 9 · 2023: 15 · 2022: 9 · 2021: 19 · 2020: 9 · 2019: 5 · 2018: 6 · 2017: 14 · 2016: 10 · 2015: 11 · 2014: 11 · 2013: 15 · 2012: 24
- FWCI
- 6.29
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- U.S. Department of Energy
- Battelle
- Japan Society for the Promotion of Science
- National Cancer Institute (R01CA079953, CA79953)
- Biological and Environmental Research
- Pacific Northwest National Laboratory
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › BRCA gene mutations in cancer
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Keywords
- BRCA1, BARD1, ubiquitin ligase, UbcH5c, UbcH7, E3 ubiquitin ligase, protein-protein binding, RING domain, tumor suppressor, ubiquitination
- MeSH terms
- Amino Acid Sequence, Binding Sites, Carrier Proteins, Cloning, Molecular, Humans, Kinetics, Ligases, Models, Molecular, Neoplasms, Magnetic Resonance Spectroscopy, Protein Conformation, Recombinant Proteins, Mutagenesis, Site-Directed, Zinc Fingers, Sequence Alignment, Sequence Homology, Amino Acid, Protein Structure, Secondary, Dimerization, BRCA1 Protein, Genes, BRCA1, Protein Subunits, Tumor Suppressor Proteins, Ubiquitin, Ubiquitin-Protein Ligases
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