FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine

Fanconi anemia (FA) is a rare hereditary disorder caused by mutations in any one of the FANC genes. FA cells are mainly characterized by extreme hypersensitivity to interstrand crosslink (ICL) agents. Additionally, the FA proteins play a crucial role in concert with homologous recombination (HR) fac...

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Autores: Peña-Gómez, María José, Moreno-Gordillo, Paula, Narmontė, Milda, García-Calderón, Clara B., Rukšėnaitė, Audronè, Klimašauskas, Saulius, Rosado, Iván V.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/282998
Acceso en línea:http://hdl.handle.net/10261/282998
Access Level:acceso abierto
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spelling FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridinePeña-Gómez, María JoséMoreno-Gordillo, PaulaNarmontė, MildaGarcía-Calderón, Clara B.Rukšėnaitė, AudronèKlimašauskas, SauliusRosado, Iván V.Fanconi anemia (FA) is a rare hereditary disorder caused by mutations in any one of the FANC genes. FA cells are mainly characterized by extreme hypersensitivity to interstrand crosslink (ICL) agents. Additionally, the FA proteins play a crucial role in concert with homologous recombination (HR) factors to protect stalled replication forks. Here, we report that the 5-methyl-2’-deoxycytidine (5mdC) demethylation (pathway) intermediate 5-hydroxymethyl-2’-deoxycytidine (5hmdC) and its deamination product 5-hydroxymethyl-2’-deoxyuridine (5hmdU) elicit a DNA damage response, chromosome aberrations, replication fork impairment and cell viability loss in the absence of FANCD2. Interestingly, replication fork instability by 5hmdC or 5hmdU was associated to the presence of Poly(ADP-ribose) polymerase 1 (PARP1) on chromatin, being both phenotypes exacerbated by olaparib treatment. Remarkably, Parp1 cells did not show any replication fork defects or sensitivity to 5hmdC or 5hmdU, suggesting that retained PARP1 at base excision repair (BER) intermediates accounts for the observed replication fork defects upon 5hmdC or 5hmdU incorporation in the absence of FANCD2. We therefore conclude that 5hmdC is deaminated in vivo to 5hmdU, whose fixation by PARP1 during BER, hinders replication fork progression and contributes to genomic instability in FA cells.The eHAP cell line were a generous gift from Dr. G. Crossan (MRC-LMB, Cambridge). DT40, NV012 and HSC72 cell lines were kindly provided by K.J. Patel (MRC WIMM, Oxford), and the MDA-MB-231, MDA-MB-436 and DLD1 BRCA2−/− were a gift from Dr. Pablo Huertas and Dr. Andrés Aguilera (CABIMER, Sevilla) respectively. We are in debt to Dr. J.A. Pérez-Simón (IBIS, Seville) for providing hosting and helpful advice. IVR´s lab is funded by MCIN/AEI/ 10.13039/501100011033, grant 18.06.03.3073 (RTI2018-100692-B-100), Consejerías de Salud, y de Economía y Conocimiento de la Junta de Andalucía, grants number PI-0005-2018, and 18.06.03.2404 (P18-RT-1271), Universidad de Sevilla “Programa Operativo FEDER Andalucía 2014–2020”, grant number 18.06.03.2319 (US-1381081), and by “ERDF A way of making Europe”. IVR is a recipient of Ramón y Cajal Contracts RYC2015-18670. SK lab was supported by ERC-2016-AdG/742654 grant.Nature Publishing GroupAgencia Estatal de Investigación (España)Ministerio de Ciencia, Innovación y Universidades (España)European CommissionJunta de AndalucíaUniversidad de SevillaEuropean Research CouncilConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2022202220222022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/282998reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-100692-B-I00info:eu-repo/grantAgreement/MINECO//RYC-2015-18670info:eu-repo/grantAgreement/EC/H2020/742654The underlying dataset has been published as supplementary material of the article in the publisher platform at 10.1038/s41419-022-04952-0http://dx.doi.org/10.1038/s41419-022-04952-0Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2829982026-05-22T06:33:51Z
dc.title.none.fl_str_mv FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
title FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
spellingShingle FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
Peña-Gómez, María José
title_short FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
title_full FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
title_fullStr FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
title_full_unstemmed FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
title_sort FANCD2 maintains replication fork stability during misincorporation of the DNA demethylation products 5-hydroxymethyl-2’-deoxycytidine and 5-hydroxymethyl-2’-deoxyuridine
dc.creator.none.fl_str_mv Peña-Gómez, María José
Moreno-Gordillo, Paula
Narmontė, Milda
García-Calderón, Clara B.
Rukšėnaitė, Audronè
Klimašauskas, Saulius
Rosado, Iván V.
author Peña-Gómez, María José
author_facet Peña-Gómez, María José
Moreno-Gordillo, Paula
Narmontė, Milda
García-Calderón, Clara B.
Rukšėnaitė, Audronè
Klimašauskas, Saulius
Rosado, Iván V.
author_role author
author2 Moreno-Gordillo, Paula
Narmontė, Milda
García-Calderón, Clara B.
Rukšėnaitė, Audronè
Klimašauskas, Saulius
Rosado, Iván V.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Agencia Estatal de Investigación (España)
Ministerio de Ciencia, Innovación y Universidades (España)
European Commission
Junta de Andalucía
Universidad de Sevilla
European Research Council
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
description Fanconi anemia (FA) is a rare hereditary disorder caused by mutations in any one of the FANC genes. FA cells are mainly characterized by extreme hypersensitivity to interstrand crosslink (ICL) agents. Additionally, the FA proteins play a crucial role in concert with homologous recombination (HR) factors to protect stalled replication forks. Here, we report that the 5-methyl-2’-deoxycytidine (5mdC) demethylation (pathway) intermediate 5-hydroxymethyl-2’-deoxycytidine (5hmdC) and its deamination product 5-hydroxymethyl-2’-deoxyuridine (5hmdU) elicit a DNA damage response, chromosome aberrations, replication fork impairment and cell viability loss in the absence of FANCD2. Interestingly, replication fork instability by 5hmdC or 5hmdU was associated to the presence of Poly(ADP-ribose) polymerase 1 (PARP1) on chromatin, being both phenotypes exacerbated by olaparib treatment. Remarkably, Parp1 cells did not show any replication fork defects or sensitivity to 5hmdC or 5hmdU, suggesting that retained PARP1 at base excision repair (BER) intermediates accounts for the observed replication fork defects upon 5hmdC or 5hmdU incorporation in the absence of FANCD2. We therefore conclude that 5hmdC is deaminated in vivo to 5hmdU, whose fixation by PARP1 during BER, hinders replication fork progression and contributes to genomic instability in FA cells.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/282998
url http://hdl.handle.net/10261/282998
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
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info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-100692-B-I00
info:eu-repo/grantAgreement/MINECO//RYC-2015-18670
info:eu-repo/grantAgreement/EC/H2020/742654
The underlying dataset has been published as supplementary material of the article in the publisher platform at 10.1038/s41419-022-04952-0
http://dx.doi.org/10.1038/s41419-022-04952-0
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
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