Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection

Ruth M Densham, Alexander J Garvin, Helen R Stone, Joanna Strachan, Robert A Baldock, Manuel Daza-Martin, Alice Fletcher, Sarah Blair-Reid, James Beesley, Balraj Johal, Laurence H Pearl, Robert Neely, Nicholas H Keep, Felicity Z Watts, Joanna R Morris

Research output: Contribution to journalArticle

Abstract

The opposing activities of 53BP1 and BRCA1 influence pathway choice in DNA double-strand-break repair. How BRCA1 counteracts the inhibitory effect of 53BP1 on DNA resection and homologous recombination is unknown. Here we identify the site of BRCA1-BARD1 required for priming ubiquitin transfer from E2∼ubiquitin and demonstrate that BRCA1-BARD1's ubiquitin ligase activity is required for repositioning 53BP1 on damaged chromatin. We confirm H2A ubiquitination by BRCA1-BARD1 and show that an H2A-ubiquitin fusion protein promotes DNA resection and repair in BARD1-deficient cells. BRCA1-BARD1's function in homologous recombination requires the chromatin remodeler SMARCAD1. SMARCAD1 binding to H2A-ubiquitin and optimal localization to sites of damage and activity in DNA repair requires its ubiquitin-binding CUE domains. SMARCAD1 is required for 53BP1 repositioning, and the need for SMARCAD1 in olaparib or camptothecin resistance is alleviated by 53BP1 loss. Thus, BRCA1-BARD1 ligase activity and subsequent SMARCAD1-dependent chromatin remodeling are critical regulators of DNA repair.

Original languageEnglish
Pages (from-to)647-55
Number of pages9
JournalNature Structural and Molecular Biology
Volume23
Issue number7
DOIs
Publication statusPublished - Jul 2016
Externally publishedYes

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Ligases
Ubiquitin
Chromatin
DNA Repair
DNA
Homologous Recombination
Camptothecin
Chromatin Assembly and Disassembly
Double-Stranded DNA Breaks
Ubiquitination
Proteins

Cite this

Densham, R. M., Garvin, A. J., Stone, H. R., Strachan, J., Baldock, R. A., Daza-Martin, M., ... Morris, J. R. (2016). Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection. Nature Structural and Molecular Biology, 23(7), 647-55. https://doi.org/10.1038/nsmb.3236
Densham, Ruth M ; Garvin, Alexander J ; Stone, Helen R ; Strachan, Joanna ; Baldock, Robert A ; Daza-Martin, Manuel ; Fletcher, Alice ; Blair-Reid, Sarah ; Beesley, James ; Johal, Balraj ; Pearl, Laurence H ; Neely, Robert ; Keep, Nicholas H ; Watts, Felicity Z ; Morris, Joanna R. / Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection. In: Nature Structural and Molecular Biology. 2016 ; Vol. 23, No. 7. pp. 647-55.
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Densham, RM, Garvin, AJ, Stone, HR, Strachan, J, Baldock, RA, Daza-Martin, M, Fletcher, A, Blair-Reid, S, Beesley, J, Johal, B, Pearl, LH, Neely, R, Keep, NH, Watts, FZ & Morris, JR 2016, 'Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection' Nature Structural and Molecular Biology, vol. 23, no. 7, pp. 647-55. https://doi.org/10.1038/nsmb.3236

Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection. / Densham, Ruth M; Garvin, Alexander J; Stone, Helen R; Strachan, Joanna; Baldock, Robert A; Daza-Martin, Manuel; Fletcher, Alice; Blair-Reid, Sarah; Beesley, James; Johal, Balraj; Pearl, Laurence H; Neely, Robert; Keep, Nicholas H; Watts, Felicity Z; Morris, Joanna R.

In: Nature Structural and Molecular Biology, Vol. 23, No. 7, 07.2016, p. 647-55.

Research output: Contribution to journalArticle

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AU - Densham, Ruth M

AU - Garvin, Alexander J

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AU - Baldock, Robert A

AU - Daza-Martin, Manuel

AU - Fletcher, Alice

AU - Blair-Reid, Sarah

AU - Beesley, James

AU - Johal, Balraj

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AU - Keep, Nicholas H

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AU - Morris, Joanna R

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N2 - The opposing activities of 53BP1 and BRCA1 influence pathway choice in DNA double-strand-break repair. How BRCA1 counteracts the inhibitory effect of 53BP1 on DNA resection and homologous recombination is unknown. Here we identify the site of BRCA1-BARD1 required for priming ubiquitin transfer from E2∼ubiquitin and demonstrate that BRCA1-BARD1's ubiquitin ligase activity is required for repositioning 53BP1 on damaged chromatin. We confirm H2A ubiquitination by BRCA1-BARD1 and show that an H2A-ubiquitin fusion protein promotes DNA resection and repair in BARD1-deficient cells. BRCA1-BARD1's function in homologous recombination requires the chromatin remodeler SMARCAD1. SMARCAD1 binding to H2A-ubiquitin and optimal localization to sites of damage and activity in DNA repair requires its ubiquitin-binding CUE domains. SMARCAD1 is required for 53BP1 repositioning, and the need for SMARCAD1 in olaparib or camptothecin resistance is alleviated by 53BP1 loss. Thus, BRCA1-BARD1 ligase activity and subsequent SMARCAD1-dependent chromatin remodeling are critical regulators of DNA repair.

AB - The opposing activities of 53BP1 and BRCA1 influence pathway choice in DNA double-strand-break repair. How BRCA1 counteracts the inhibitory effect of 53BP1 on DNA resection and homologous recombination is unknown. Here we identify the site of BRCA1-BARD1 required for priming ubiquitin transfer from E2∼ubiquitin and demonstrate that BRCA1-BARD1's ubiquitin ligase activity is required for repositioning 53BP1 on damaged chromatin. We confirm H2A ubiquitination by BRCA1-BARD1 and show that an H2A-ubiquitin fusion protein promotes DNA resection and repair in BARD1-deficient cells. BRCA1-BARD1's function in homologous recombination requires the chromatin remodeler SMARCAD1. SMARCAD1 binding to H2A-ubiquitin and optimal localization to sites of damage and activity in DNA repair requires its ubiquitin-binding CUE domains. SMARCAD1 is required for 53BP1 repositioning, and the need for SMARCAD1 in olaparib or camptothecin resistance is alleviated by 53BP1 loss. Thus, BRCA1-BARD1 ligase activity and subsequent SMARCAD1-dependent chromatin remodeling are critical regulators of DNA repair.

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SN - 1545-9993

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