Small-Molecule Disruption of RAD52 Rings as a Mechanism for Precision Medicine in BRCA-Deficient Cancers

Gurushankar Chandramouly; B McDevitt; Katherine Sullivan; Tatiana Kent; Antonio Luz; J. Fraser Glickman; Mark Andrake; Tomasz Skorski; Richard T. Pomerantz

doi:10.1016/j.chembiol.2015.10.003

Small-Molecule Disruption of RAD52 Rings as a Mechanism for Precision Medicine in BRCA-Deficient Cancers

Gurushankar Chandramouly, B McDevitt, Katherine Sullivan, Tatiana Kent, Antonio Luz, J. Fraser Glickman, Mark Andrake, Tomasz Skorski, Richard T. Pomerantz

Research output: Contribution to journal › Article › peer-review

68 Scopus citations

Abstract

Summary Suppression of RAD52 causes synthetic lethality in BRCA-deficient cells. Yet pharmacological inhibition of RAD52, which binds single-strand DNA (ssDNA) and lacks enzymatic activity, has not been demonstrated. Here, we identify the small molecule 6-hydroxy-DL-dopa (6-OH-dopa) as a major allosteric inhibitor of the RAD52 ssDNA binding domain. For example, we find that multiple small molecules bind to and completely transform RAD52 undecamer rings into dimers, which abolishes the ssDNA binding channel observed in crystal structures. 6-OH-Dopa also disrupts RAD52 heptamer and undecamer ring superstructures, and suppresses RAD52 recruitment and recombination activity in cells with negligible effects on other double-strand break repair pathways. Importantly, we show that 6-OH-dopa selectively inhibits the proliferation of BRCA-deficient cancer cells, including those obtained from leukemia patients. Taken together, these data demonstrate small-molecule disruption of RAD52 rings as a promising mechanism for precision medicine in BRCA-deficient cancers.

Original language	English
Pages (from-to)	1491-1504
Number of pages	14
Journal	Chemistry and Biology
Volume	22
Issue number	11
DOIs	https://doi.org/10.1016/j.chembiol.2015.10.003
State	Published - Nov 19 2015

Keywords

Allosteric Regulation
Apoptosis/drug effects
BRCA1 Protein/deficiency
BRCA2 Protein/deficiency
Cell Line
Cell Proliferation/drug effects
DNA Damage/drug effects
DNA, Single-Stranded/chemistry
Dihydroxyphenylalanine/analogs & derivatives
Electrophoretic Mobility Shift Assay
Humans
Inhibitory Concentration 50
Microscopy, Fluorescence
Neoplasms/genetics
Protein Binding
RNA Interference
RNA, Small Interfering/metabolism
Rad52 DNA Repair and Recombination Protein/antagonists & inhibitors
Recombinant Proteins/biosynthesis
Small Molecule Libraries/chemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.chembiol.2015.10.003

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title = "Small-Molecule Disruption of RAD52 Rings as a Mechanism for Precision Medicine in BRCA-Deficient Cancers",

abstract = "Summary Suppression of RAD52 causes synthetic lethality in BRCA-deficient cells. Yet pharmacological inhibition of RAD52, which binds single-strand DNA (ssDNA) and lacks enzymatic activity, has not been demonstrated. Here, we identify the small molecule 6-hydroxy-DL-dopa (6-OH-dopa) as a major allosteric inhibitor of the RAD52 ssDNA binding domain. For example, we find that multiple small molecules bind to and completely transform RAD52 undecamer rings into dimers, which abolishes the ssDNA binding channel observed in crystal structures. 6-OH-Dopa also disrupts RAD52 heptamer and undecamer ring superstructures, and suppresses RAD52 recruitment and recombination activity in cells with negligible effects on other double-strand break repair pathways. Importantly, we show that 6-OH-dopa selectively inhibits the proliferation of BRCA-deficient cancer cells, including those obtained from leukemia patients. Taken together, these data demonstrate small-molecule disruption of RAD52 rings as a promising mechanism for precision medicine in BRCA-deficient cancers.",

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author = "Gurushankar Chandramouly and B McDevitt and Katherine Sullivan and Tatiana Kent and Antonio Luz and Glickman, {J. Fraser} and Mark Andrake and Tomasz Skorski and Pomerantz, {Richard T.}",

note = "Publisher Copyright: {\textcopyright} 2015 Elsevier Ltd.",

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doi = "10.1016/j.chembiol.2015.10.003",

language = "English",

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T1 - Small-Molecule Disruption of RAD52 Rings as a Mechanism for Precision Medicine in BRCA-Deficient Cancers

AU - Chandramouly, Gurushankar

AU - McDevitt, B

AU - Sullivan, Katherine

AU - Kent, Tatiana

AU - Luz, Antonio

AU - Glickman, J. Fraser

AU - Andrake, Mark

AU - Skorski, Tomasz

AU - Pomerantz, Richard T.

PY - 2015/11/19

Y1 - 2015/11/19

N2 - Summary Suppression of RAD52 causes synthetic lethality in BRCA-deficient cells. Yet pharmacological inhibition of RAD52, which binds single-strand DNA (ssDNA) and lacks enzymatic activity, has not been demonstrated. Here, we identify the small molecule 6-hydroxy-DL-dopa (6-OH-dopa) as a major allosteric inhibitor of the RAD52 ssDNA binding domain. For example, we find that multiple small molecules bind to and completely transform RAD52 undecamer rings into dimers, which abolishes the ssDNA binding channel observed in crystal structures. 6-OH-Dopa also disrupts RAD52 heptamer and undecamer ring superstructures, and suppresses RAD52 recruitment and recombination activity in cells with negligible effects on other double-strand break repair pathways. Importantly, we show that 6-OH-dopa selectively inhibits the proliferation of BRCA-deficient cancer cells, including those obtained from leukemia patients. Taken together, these data demonstrate small-molecule disruption of RAD52 rings as a promising mechanism for precision medicine in BRCA-deficient cancers.

AB - Summary Suppression of RAD52 causes synthetic lethality in BRCA-deficient cells. Yet pharmacological inhibition of RAD52, which binds single-strand DNA (ssDNA) and lacks enzymatic activity, has not been demonstrated. Here, we identify the small molecule 6-hydroxy-DL-dopa (6-OH-dopa) as a major allosteric inhibitor of the RAD52 ssDNA binding domain. For example, we find that multiple small molecules bind to and completely transform RAD52 undecamer rings into dimers, which abolishes the ssDNA binding channel observed in crystal structures. 6-OH-Dopa also disrupts RAD52 heptamer and undecamer ring superstructures, and suppresses RAD52 recruitment and recombination activity in cells with negligible effects on other double-strand break repair pathways. Importantly, we show that 6-OH-dopa selectively inhibits the proliferation of BRCA-deficient cancer cells, including those obtained from leukemia patients. Taken together, these data demonstrate small-molecule disruption of RAD52 rings as a promising mechanism for precision medicine in BRCA-deficient cancers.

KW - Allosteric Regulation

KW - Apoptosis/drug effects

KW - BRCA1 Protein/deficiency

KW - BRCA2 Protein/deficiency

KW - Cell Line

KW - Cell Proliferation/drug effects

KW - DNA Damage/drug effects

KW - DNA, Single-Stranded/chemistry

KW - Dihydroxyphenylalanine/analogs & derivatives

KW - Electrophoretic Mobility Shift Assay

KW - Humans

KW - Inhibitory Concentration 50

KW - Microscopy, Fluorescence

KW - Neoplasms/genetics

KW - Protein Binding

KW - RNA Interference

KW - RNA, Small Interfering/metabolism

KW - Rad52 DNA Repair and Recombination Protein/antagonists & inhibitors

KW - Recombinant Proteins/biosynthesis

KW - Small Molecule Libraries/chemistry

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DO - 10.1016/j.chembiol.2015.10.003

M3 - Article

C2 - 26548611

SN - 1074-5521

VL - 22

SP - 1491

EP - 1504

JO - Chemistry and Biology

JF - Chemistry and Biology

IS - 11

ER -

Small-Molecule Disruption of RAD52 Rings as a Mechanism for Precision Medicine in BRCA-Deficient Cancers

Abstract

Keywords

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