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)	647-655
Number of pages	9
Journal	Nature Structural & Molecular Biology
Volume	23
Issue number	7
Early online date	30 May 2016
DOIs	https://doi.org/10.1038/nsmb.3236
Publication status	Published - 1 Jul 2016

Keywords

BRCA1 Protein/genetics
Binding Sites
Camptothecin/pharmacology
Chromatin/chemistry
Cloning, Molecular
DNA Breaks, Double-Stranded
DNA Cleavage/drug effects
DNA Helicases/genetics
DNA, Neoplasm/genetics
Escherichia coli/genetics
Gene Expression
Gene Expression Regulation, Neoplastic
HeLa Cells
Histones/genetics
Humans
Models, Molecular
Phthalazines/pharmacology
Piperazines/pharmacology
Protein Binding
Recombinant Proteins/genetics
Recombinational DNA Repair
Signal Transduction
Tumor Suppressor Proteins/genetics
Tumor Suppressor p53-Binding Protein 1/genetics
Ubiquitin/genetics
Ubiquitin-Protein Ligases/genetics
Ubiquitination/drug effects

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10.1038/nsmb.3236

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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 & Molecular Biology, 23(7), 647-655. 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.",

keywords = "BRCA1 Protein/genetics, Binding Sites, Camptothecin/pharmacology, Chromatin/chemistry, Cloning, Molecular, DNA Breaks, Double-Stranded, DNA Cleavage/drug effects, DNA Helicases/genetics, DNA, Neoplasm/genetics, Escherichia coli/genetics, Gene Expression, Gene Expression Regulation, Neoplastic, HeLa Cells, Histones/genetics, Humans, Models, Molecular, Phthalazines/pharmacology, Piperazines/pharmacology, Protein Binding, Recombinant Proteins/genetics, Recombinational DNA Repair, Signal Transduction, Tumor Suppressor Proteins/genetics, Tumor Suppressor p53-Binding Protein 1/genetics, Ubiquitin/genetics, Ubiquitin-Protein Ligases/genetics, Ubiquitination/drug effects",

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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doi = "10.1038/nsmb.3236",

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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 & Molecular Biology, vol. 23, no. 7, pp. 647-655. https://doi.org/10.1038/nsmb.3236

TY - JOUR

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 - 2016/7/1

Y1 - 2016/7/1

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.

KW - BRCA1 Protein/genetics

KW - Binding Sites

KW - Camptothecin/pharmacology

KW - Chromatin/chemistry

KW - Cloning, Molecular

KW - DNA Breaks, Double-Stranded

KW - DNA Cleavage/drug effects

KW - DNA Helicases/genetics

KW - DNA, Neoplasm/genetics

KW - Escherichia coli/genetics

KW - Gene Expression

KW - Gene Expression Regulation, Neoplastic

KW - HeLa Cells

KW - Histones/genetics

KW - Humans

KW - Models, Molecular

KW - Phthalazines/pharmacology

KW - Piperazines/pharmacology

KW - Protein Binding

KW - Recombinant Proteins/genetics

KW - Recombinational DNA Repair

KW - Signal Transduction

KW - Tumor Suppressor Proteins/genetics

KW - Tumor Suppressor p53-Binding Protein 1/genetics

KW - Ubiquitin/genetics

KW - Ubiquitin-Protein Ligases/genetics

KW - Ubiquitination/drug effects

UR - https://sro.sussex.ac.uk/

U2 - 10.1038/nsmb.3236

DO - 10.1038/nsmb.3236

M3 - Article

C2 - 27239795

SN - 1545-9993

VL - 23

SP - 647

EP - 655

JO - Nature Structural & Molecular Biology

JF - Nature Structural & Molecular Biology

IS - 7

ER -

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

Abstract

Keywords

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