Optimised oligonucleotide substrates to assay XPF-ERCC1 nuclease activity for the discovery of DNA repair inhibitors

Adam M Thomas; Sanja Brolih; Joanna F McGouran; Afaf H El-Sagheer; Denis Ptchelkine; Morgan Jones; Neil Q McDonald; Peter J McHugh; Tom Brown

doi:10.1039/c9cc05476f

Optimised oligonucleotide substrates to assay XPF-ERCC1 nuclease activity for the discovery of DNA repair inhibitors

Chem Commun (Camb). 2019 Sep 26;55(78):11671-11674. doi: 10.1039/c9cc05476f.

Authors

Adam M Thomas¹, Sanja Brolih², Joanna F McGouran³, Afaf H El-Sagheer⁴, Denis Ptchelkine², Morgan Jones⁵, Neil Q McDonald⁶, Peter J McHugh², Tom Brown³

Affiliations

¹ Department of Chemistry, University of Oxford, Chemistry Research Laboratory, 12 Mansfield Road, Oxford, OX1 3TA, UK. tom.brown@chem.ox.ac.uk and Department of Oncology, MRC-Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DS, UK and The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
² Department of Oncology, MRC-Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DS, UK.
³ Department of Chemistry, University of Oxford, Chemistry Research Laboratory, 12 Mansfield Road, Oxford, OX1 3TA, UK. tom.brown@chem.ox.ac.uk.
⁴ Department of Chemistry, University of Oxford, Chemistry Research Laboratory, 12 Mansfield Road, Oxford, OX1 3TA, UK. tom.brown@chem.ox.ac.uk and Chemistry Branch, Department of Science and Mathematics, Faculty of Petroleum and Mining Engineering, Suez University, Suez 43721, Egypt.
⁵ The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
⁶ The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK and Institute of Structural and Molecular Biology, Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, UK.

PMID: 31497827
DOI: 10.1039/c9cc05476f

Abstract

We report the design and optimisation of novel oligonucleotide substrates for a sensitive fluorescence assay for high-throughput screening and functional studies of the DNA repair enzyme, XPF-ERCC1, with a view to accelerating inhibitor and drug discovery.

MeSH terms

DNA Repair*
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Dimerization
Endonucleases / chemistry
Endonucleases / genetics
Endonucleases / metabolism*
Humans
Oligonucleotides / chemistry
Oligonucleotides / metabolism
Substrate Specificity
Temperature

Substances

DNA-Binding Proteins
Oligonucleotides
Endonucleases

Grants and funding

MR/R009368/1/MRC_/Medical Research Council/United Kingdom