CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions
| Autor: | Andrea McEwan, Megha Chandrashekhar, Olga Murina, Maryou B. Lambros, Valerie G. Brunton, Daniel Durocher, Angelo Agathanggelou, Martin A M Reijns, Jason Moffat, Tatjana Stankovic, Michal Zimmermann, Stephane Angers, Traver Hart, Adeline Fluteau, Michael Aregger, Paul Moss, Morwenna Muir, Wei Yuan, Rachel C. Challis, Matthew Clarke, Andrew P. Jackson, Shankara Paneesha, Johann S. de Bono, Žygimantė Tarnauskaitė |
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| Rok vydání: | 2018 |
| Předmět: |
DNA Replication
Male 0301 basic medicine Genome instability DNA Repair DNA repair Ribonucleotide excision repair Ribonuclease H Genes BRCA1 Poly (ADP-Ribose) Polymerase-1 Poly(ADP-ribose) Polymerase Inhibitors Biology Poly (ADP-Ribose) Polymerase Inhibitor Piperazines Cell Line Olaparib Mice 03 medical and health sciences chemistry.chemical_compound PARP1 Neoplasms Animals Humans CRISPR Gene Editing Genome Multidisciplinary BRCA1 Protein Prostatic Neoplasms Ribonucleotides Leukemia Lymphocytic Chronic B-Cell Xenograft Model Antitumor Assays 3. Good health 030104 developmental biology DNA Topoisomerases Type I chemistry Cancer research Phthalazines Female CRISPR-Cas Systems Synthetic Lethal Mutations Homologous recombination DNA Damage HeLa Cells |
| Zdroj: | Zimmermann, M, Murina, O, Reijns, M A M, Agathanggelou, A, Challis, R, Tarnauskaite, Z, Muir, M, Fluteau, A, Aregger, M, McEwan, A, Yuan, W, Clarke, M, Lambros, M B, Paneesha, S, Moss, P, Chandrashekhar, M, Angers, S, Moffat, J, Brunton, V G, Hart, T, de Bono, J, Stankovic, T, Jackson, A P & Durocher, D 2019, ' CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions ', Nature, vol. 559, no. 7713, pp. 285–289 . https://doi.org/10.1038/s41586-018-0291-z |
| ISSN: | 1476-4687 0028-0836 |
| DOI: | 10.1038/s41586-018-0291-z |
| Popis: | The observation that BRCA1- and BRCA2-deficient cells are sensitive to inhibitors of poly(ADP–ribose) polymerase (PARP) has spurred the development of cancer therapies that use these inhibitors to target deficiencies in homologous recombination. The cytotoxicity of PARP inhibitors depends on PARP trapping, the formation of non-covalent protein–DNA adducts composed of inhibited PARP1 bound to DNA lesions of unclear origins. To address the nature of such lesions and the cellular consequences of PARP trapping, we undertook three CRISPR (clustered regularly interspersed palindromic repeats) screens to identify genes and pathways that mediate cellular resistance to olaparib, a clinically approved PARP inhibitor. Here we present a high-confidence set of 73 genes, which when mutated cause increased sensitivity to PARP inhibitors. In addition to an expected enrichment for genes related to homologous recombination, we discovered that mutations in all three genes encoding ribonuclease H2 sensitized cells to PARP inhibition. We establish that the underlying cause of the PARP-inhibitor hypersensitivity of cells deficient in ribonuclease H2 is impaired ribonucleotide excision repair. Embedded ribonucleotides, which are abundant in the genome of cells deficient in ribonucleotide excision repair, are substrates for cleavage by topoisomerase 1, resulting in PARP-trapping lesions that impede DNA replication and endanger genome integrity. We conclude that genomic ribonucleotides are a hitherto unappreciated source of PARP-trapping DNA lesions, and that the frequent deletion of RNASEH2B in metastatic prostate cancer and chronic lymphocytic leukaemia could provide an opportunity to exploit these findings therapeutically. |
| Databáze: | OpenAIRE |
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