XRCC4 protein interactions with XRCC4-like factor (XLF) create an extended grooved scaffold for DNA ligation and double strand break repair
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The XRCC4-like factor (XLF)-XRCC4 complex is essential for nonhomologous end joining, the major repair pathway for DNA double strand breaks in human cells. Yet, how XLF binds XRCC4 and impacts nonhomologous end joining functions has been enigmatic. Here, we report the XLF-XRCC4 complex crystal structure in combination with biophysical and mutational analyses to define the XLF-XRCC4 interactions. Crystal and solution structures plus mutations characterize alternating XRCC4 and XLF head domain interfaces forming parallel super-helical filaments. XLF Leu-115 ("Leu-lock") inserts into a hydrophobic pocket formed by XRCC4 Met-59, Met-61, Lys-65, Lys-99, Phe-106, and Leu-108 in synergy with pseudo-symmetric β-zipper hydrogen bonds to drive specificity. XLF C terminus and DNA enhance parallel filament formation. Super-helical XLF-XRCC4 filaments form a positively charged channel to bind DNA and align ends for efficient ligation. Collective results reveal how human XLF and XRCC4 interact to bind DNA, suggest consequences of patient mutations, and support a unified molecular mechanism for XLF-XRCC4 stimulation of DNA ligation.
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XLF/Cernunnos joins with XRCC4 to align DNA termini for more efficient end-joining.
“The XRCC4 Like Factor (XLF/Cernunnos) functions biochemically to join with the XRCC4 protein to align DNA termini for more efficient end-joining [66,67].”
From Integration of DNA damage and repair with murine double-minute 2 (Mdm2) in tumorigenesis (2012) · cites this paper as [67]
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- Authors
- Hammel, M., Rey, M., Yu, Y., Mani, R. S., Classen, S., Liu, M., Pique, M. E., Fang, S., Mahaney, B. L., Weinfeld, M., Schriemer, D. C., Lees-Miller, S. P., & Tainer, J. A.
- Affiliations
- Lawrence Berkeley National Laboratory (US)
- University of Calgary (CA)
- University of Alberta (CA)
- Scripps Research Institute (US)
- Corresponding author
- Hammel, Michal
- Tainer, John A.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2011-09
- Volume
- 286
- Issue
- 37
- Pages
- 32638-32650
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC3173232
- OpenAlex
- W2139045592
- Cited by
- 179
- References
- 52
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 3 · 2023: 10 · 2022: 6 · 2021: 16 · 2020: 11 · 2019: 17 · 2018: 9 · 2017: 18 · 2016: 15 · 2015: 15 · 2014: 18 · 2013: 18 · 2012: 17
- FWCI
- 7.02
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Libraries and Cultural Resources (University of Calgary) (submittedVersion)
- Funders
- U.S. Department of Energy (-AC02-05CH11231, 05CH11231, DE AC02 05CH11231, DE-AC02-)
- Alberta Cancer Foundation
- University of Alberta
- National Institutes of Health (DE-AC02–05CH11231, CA92584, 69139)
- Canadian Institutes of Health Research (DE-AC02-05CH11231)
- National Cancer Institute (P01-CA 92584, P01CA092584)
- Lawrence Berkeley National Laboratory (AC02‐05CH11231, DE-AC02-05CH11231, 05CH11231)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- XRCC4, X-ray luminosity function, non-homologous end joining, DNA double-strand break repair, DNA ligation, protein-protein interaction, crystal structure, head domain, DNA binding
- MeSH terms
- Cell Line, DNA, DNA Repair, DNA-Binding Proteins, Humans, Protein Binding, Protein Structure, Secondary, Crystallography, X-Ray, Protein Structure, Quaternary, DNA Repair Enzymes, DNA Breaks, Double-Stranded
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