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

doi:10.1038/nsmb.3236

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 journal › Article › peer-review

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 language	English
Pages (from-to)	647-55
Number of pages	9
Journal	Nature Structural and Molecular Biology
Volume	23
Issue number	7
DOIs	https://doi.org/10.1038/nsmb.3236
Publication status	Published - 30 May 2016
Externally published	Yes

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https://research.birmingham.ac.uk/portal/en/publications/human-brca1bard1-ubiquitin-ligase-activity-counteracts-chromatin-barriers-to-dna-resection(bbb5e8ad-ac69-48d0-93b9-fc158e95f903).html

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Densham, R. M., Garvin, A. J., Stone, H. R., Strachan, J., Baldock, R. A., Daza-Martin, M., Fletcher, A., Blair-Reid, S., Beesley, J., Johal, B., Pearl, L. H., Neely, R., Keep, N. H., Watts, F. Z., & 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

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title = "Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection",

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.",

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

year = "2016",

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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

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T1 - Human BRCA1-BARD1 ubiquitin ligase activity counteracts chromatin barriers to DNA resection

AU - Densham, Ruth M

AU - Garvin, Alexander J

AU - Stone, Helen R

AU - Strachan, Joanna

AU - Baldock, Robert A

AU - Daza-Martin, Manuel

AU - Fletcher, Alice

AU - Blair-Reid, Sarah

AU - Beesley, James

AU - Johal, Balraj

AU - Pearl, Laurence H

AU - Neely, Robert

AU - Keep, Nicholas H

AU - Watts, Felicity Z

AU - Morris, Joanna R

PY - 2016/5/30

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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.

U2 - 10.1038/nsmb.3236

DO - 10.1038/nsmb.3236

M3 - Article

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VL - 23

SP - 647

EP - 655

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 7

ER -

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

Abstract

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