Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells

Autores
Chatgilialoglu, Chryssostomos; Krokidis, Marios G.; Masi, Annalisa; Barata Vallejo, Sebastian; Ferreri, Carla; Pascucci, Barbara; D?Errico, Mariarosaria
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Mitochondrial (mt) DNA and nuclear (n) DNA have known structures and roles in cells; however, they are rarely compared under specific conditions such as oxidative or degenerative environments that can create damage to the DNA base moieties. Six purine lesions were ascertained in the mtDNA of wild type (wt) CSA (CS3BE–wtCSA) and wtCSB (CS1AN–wtCSB) cells and defective counterparts CS3BE and CS1AN in comparison with the corresponding total (t) DNA (t = n + mt). In particular, the four 5′,8–cyclopurine (cPu) and the two 8–oxo–purine (8–oxo–Pu) lesions were accurately quantified by LC–MS/MS analysis using isotopomeric internal standards after an enzymatic digestion procedure. The 8–oxo–Pu levels were found to be in the range of 25–50 lesions/107 nucleotides in both the mtDNA and tDNA. The four cPu were undetectable in the mtDNA both in defective cells and in the wt counterparts (CSA and CSB), contrary to their detection in tDNA, indicating a nonappearance of hydroxyl radical (HO•) reactivity within the mtDNA. In order to assess the HO• reactivity towards purine nucleobases in the two genetic materials, we performed γ–radiolysis experiments coupled with the 8–oxo–Pu and cPu quantifications on isolated mtDNA and tDNA from wtCSB cells. In the latter experiments, all six purine lesions were detected in both of the DNA, showing a higher resistance to HO• attack in the case of mtDNA compared with tDNA, likely due to their different DNA helical topology influencing the relative abundance of the lesions.
Fil: Chatgilialoglu, Chryssostomos. Adam Mickiewicz University; Polonia
Fil: Krokidis, Marios G.. No especifíca;
Fil: Masi, Annalisa. Consiglio Nazionale delle Ricerche; Italia
Fil: Barata Vallejo, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Ferreri, Carla. Istituto Per la Sintesi Organica E la Fotoreattivita; Italia
Fil: Pascucci, Barbara. Consiglio Nazionale delle Ricerche; Italia
Fil: D?Errico, Mariarosaria. Istituto Superiore di Sanità; Italia
Materia
5′,8–CYCLOPURINES
8–OXO–DG
COCKAYNE SYNDROME
GAMMA RADIOLYSIS
HYDROXYL RADICAL
ISOTOPE DILUTION LC–MS/MS
MITOCHONDRIAL AND NUCLEAR DNA DAMAGE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/217032
Acceder
id CONICETDig_3d6c2aa8f2b1775afcd35f4aff259a2b
oai_identifier_str oai:ri.conicet.gov.ar:11336/217032
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome CellsChatgilialoglu, ChryssostomosKrokidis, Marios G.Masi, AnnalisaBarata Vallejo, SebastianFerreri, CarlaPascucci, BarbaraD?Errico, Mariarosaria5′,8–CYCLOPURINES8–OXO–DGCOCKAYNE SYNDROMEGAMMA RADIOLYSISHYDROXYL RADICALISOTOPE DILUTION LC–MS/MSMITOCHONDRIAL AND NUCLEAR DNA DAMAGEhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Mitochondrial (mt) DNA and nuclear (n) DNA have known structures and roles in cells; however, they are rarely compared under specific conditions such as oxidative or degenerative environments that can create damage to the DNA base moieties. Six purine lesions were ascertained in the mtDNA of wild type (wt) CSA (CS3BE–wtCSA) and wtCSB (CS1AN–wtCSB) cells and defective counterparts CS3BE and CS1AN in comparison with the corresponding total (t) DNA (t = n + mt). In particular, the four 5′,8–cyclopurine (cPu) and the two 8–oxo–purine (8–oxo–Pu) lesions were accurately quantified by LC–MS/MS analysis using isotopomeric internal standards after an enzymatic digestion procedure. The 8–oxo–Pu levels were found to be in the range of 25–50 lesions/107 nucleotides in both the mtDNA and tDNA. The four cPu were undetectable in the mtDNA both in defective cells and in the wt counterparts (CSA and CSB), contrary to their detection in tDNA, indicating a nonappearance of hydroxyl radical (HO•) reactivity within the mtDNA. In order to assess the HO• reactivity towards purine nucleobases in the two genetic materials, we performed γ–radiolysis experiments coupled with the 8–oxo–Pu and cPu quantifications on isolated mtDNA and tDNA from wtCSB cells. In the latter experiments, all six purine lesions were detected in both of the DNA, showing a higher resistance to HO• attack in the case of mtDNA compared with tDNA, likely due to their different DNA helical topology influencing the relative abundance of the lesions.Fil: Chatgilialoglu, Chryssostomos. Adam Mickiewicz University; PoloniaFil: Krokidis, Marios G.. No especifíca;Fil: Masi, Annalisa. Consiglio Nazionale delle Ricerche; ItaliaFil: Barata Vallejo, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Ferreri, Carla. Istituto Per la Sintesi Organica E la Fotoreattivita; ItaliaFil: Pascucci, Barbara. Consiglio Nazionale delle Ricerche; ItaliaFil: D?Errico, Mariarosaria. Istituto Superiore di Sanità; ItaliaMDPI2022-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/217032Chatgilialoglu, Chryssostomos; Krokidis, Marios G.; Masi, Annalisa; Barata Vallejo, Sebastian; Ferreri, Carla; et al.; Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells; MDPI; Biomolecules; 12; 11; 11-2022; 1-142218-273XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.3390/biom12111630info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:06:40Zoai:ri.conicet.gov.ar:11336/217032instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:06:41.185CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
title Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
spellingShingle Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
Chatgilialoglu, Chryssostomos
5′,8–CYCLOPURINES
8–OXO–DG
COCKAYNE SYNDROME
GAMMA RADIOLYSIS
HYDROXYL RADICAL
ISOTOPE DILUTION LC–MS/MS
MITOCHONDRIAL AND NUCLEAR DNA DAMAGE
title_short Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
title_full Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
title_fullStr Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
title_full_unstemmed Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
title_sort Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells
dc.creator.none.fl_str_mv Chatgilialoglu, Chryssostomos
Krokidis, Marios G.
Masi, Annalisa
Barata Vallejo, Sebastian
Ferreri, Carla
Pascucci, Barbara
D?Errico, Mariarosaria
author Chatgilialoglu, Chryssostomos
author_facet Chatgilialoglu, Chryssostomos
Krokidis, Marios G.
Masi, Annalisa
Barata Vallejo, Sebastian
Ferreri, Carla
Pascucci, Barbara
D?Errico, Mariarosaria
author_role author
author2 Krokidis, Marios G.
Masi, Annalisa
Barata Vallejo, Sebastian
Ferreri, Carla
Pascucci, Barbara
D?Errico, Mariarosaria
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv 5′,8–CYCLOPURINES
8–OXO–DG
COCKAYNE SYNDROME
GAMMA RADIOLYSIS
HYDROXYL RADICAL
ISOTOPE DILUTION LC–MS/MS
MITOCHONDRIAL AND NUCLEAR DNA DAMAGE
topic 5′,8–CYCLOPURINES
8–OXO–DG
COCKAYNE SYNDROME
GAMMA RADIOLYSIS
HYDROXYL RADICAL
ISOTOPE DILUTION LC–MS/MS
MITOCHONDRIAL AND NUCLEAR DNA DAMAGE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Mitochondrial (mt) DNA and nuclear (n) DNA have known structures and roles in cells; however, they are rarely compared under specific conditions such as oxidative or degenerative environments that can create damage to the DNA base moieties. Six purine lesions were ascertained in the mtDNA of wild type (wt) CSA (CS3BE–wtCSA) and wtCSB (CS1AN–wtCSB) cells and defective counterparts CS3BE and CS1AN in comparison with the corresponding total (t) DNA (t = n + mt). In particular, the four 5′,8–cyclopurine (cPu) and the two 8–oxo–purine (8–oxo–Pu) lesions were accurately quantified by LC–MS/MS analysis using isotopomeric internal standards after an enzymatic digestion procedure. The 8–oxo–Pu levels were found to be in the range of 25–50 lesions/107 nucleotides in both the mtDNA and tDNA. The four cPu were undetectable in the mtDNA both in defective cells and in the wt counterparts (CSA and CSB), contrary to their detection in tDNA, indicating a nonappearance of hydroxyl radical (HO•) reactivity within the mtDNA. In order to assess the HO• reactivity towards purine nucleobases in the two genetic materials, we performed γ–radiolysis experiments coupled with the 8–oxo–Pu and cPu quantifications on isolated mtDNA and tDNA from wtCSB cells. In the latter experiments, all six purine lesions were detected in both of the DNA, showing a higher resistance to HO• attack in the case of mtDNA compared with tDNA, likely due to their different DNA helical topology influencing the relative abundance of the lesions.
Fil: Chatgilialoglu, Chryssostomos. Adam Mickiewicz University; Polonia
Fil: Krokidis, Marios G.. No especifíca;
Fil: Masi, Annalisa. Consiglio Nazionale delle Ricerche; Italia
Fil: Barata Vallejo, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Ferreri, Carla. Istituto Per la Sintesi Organica E la Fotoreattivita; Italia
Fil: Pascucci, Barbara. Consiglio Nazionale delle Ricerche; Italia
Fil: D?Errico, Mariarosaria. Istituto Superiore di Sanità; Italia
description Mitochondrial (mt) DNA and nuclear (n) DNA have known structures and roles in cells; however, they are rarely compared under specific conditions such as oxidative or degenerative environments that can create damage to the DNA base moieties. Six purine lesions were ascertained in the mtDNA of wild type (wt) CSA (CS3BE–wtCSA) and wtCSB (CS1AN–wtCSB) cells and defective counterparts CS3BE and CS1AN in comparison with the corresponding total (t) DNA (t = n + mt). In particular, the four 5′,8–cyclopurine (cPu) and the two 8–oxo–purine (8–oxo–Pu) lesions were accurately quantified by LC–MS/MS analysis using isotopomeric internal standards after an enzymatic digestion procedure. The 8–oxo–Pu levels were found to be in the range of 25–50 lesions/107 nucleotides in both the mtDNA and tDNA. The four cPu were undetectable in the mtDNA both in defective cells and in the wt counterparts (CSA and CSB), contrary to their detection in tDNA, indicating a nonappearance of hydroxyl radical (HO•) reactivity within the mtDNA. In order to assess the HO• reactivity towards purine nucleobases in the two genetic materials, we performed γ–radiolysis experiments coupled with the 8–oxo–Pu and cPu quantifications on isolated mtDNA and tDNA from wtCSB cells. In the latter experiments, all six purine lesions were detected in both of the DNA, showing a higher resistance to HO• attack in the case of mtDNA compared with tDNA, likely due to their different DNA helical topology influencing the relative abundance of the lesions.
publishDate 2022
dc.date.none.fl_str_mv 2022-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/217032
Chatgilialoglu, Chryssostomos; Krokidis, Marios G.; Masi, Annalisa; Barata Vallejo, Sebastian; Ferreri, Carla; et al.; Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells; MDPI; Biomolecules; 12; 11; 11-2022; 1-14
2218-273X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/217032
identifier_str_mv Chatgilialoglu, Chryssostomos; Krokidis, Marios G.; Masi, Annalisa; Barata Vallejo, Sebastian; Ferreri, Carla; et al.; Assessing the Formation of Purine Lesions in Mitochondrial DNA of Cockayne Syndrome Cells; MDPI; Biomolecules; 12; 11; 11-2022; 1-14
2218-273X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.3390/biom12111630
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.069144

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