Both replication bypass fidelity and repair efficiency influence the yield of mutations per target dose in intact mammalian cells induced by benzo[a]pyrene-diol-epoxide and dibenzo[a,l]pyrene-diol-epoxide.

2.50
Hdl Handle:
http://hdl.handle.net/10146/77381
Title:
Both replication bypass fidelity and repair efficiency influence the yield of mutations per target dose in intact mammalian cells induced by benzo[a]pyrene-diol-epoxide and dibenzo[a,l]pyrene-diol-epoxide.
Authors:
Lagerqvist, Anne; Hakansson, Daniel; Prochazka, Gabriela; Lundin, Cecilia; Dreij, Kristian; Segerback, Dan; Jernstrom, Bengt; Tornqvist, Margareta; Seidel, Albrecht; Erixon, Klaus; Jenssen, Dag
Abstract:
Mutations induced by polycyclic aromatic hydrocarbons (PAH) are expected to be produced when error-prone DNA replication occurs across unrepaired DNA lesions formed by reactive PAH metabolites such as diol epoxides. The mutagenicity of the two PAH-diol epoxides (+)-anti-7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) and (+/-)-anti-11,12-dihydroxy-13,14-epoxy-11,12,13,14-tetrahydrodibenzo[a,l]pyrene (DBPDE) was compared in nucleotide excision repair (NER) proficient and deficient hamster cell lines. We applied the (32)P-postlabelling assay to analyze adduct levels and the hprt gene mutation assay for monitoring mutations. It was found that the mutagenicity per target dose was 4 times higher for DBPDE compared to BPDE in NER proficient cells while in NER deficient cells, the mutagenicity per target dose was 1.4 times higher for BPDE. In order to investigate to what extent the mutagenicity of the different adducts in NER proficient cells was influenced by repair or replication bypass, we measured the overall NER incision rate, the rate of adduct removal, the rate of replication bypass and the frequency of induced recombination in the hprt gene. The results suggest that NER of BPDE lesions are 5 times more efficient than for DBPDE lesions, in NER proficient cells. However, DBPDE adducts block replication more efficiently and also induce 6 times more recombination events in the hprt gene than adducts of BPDE, suggesting that DBPDE adducts are, to a larger extent, bypassed by homologous recombination. The results obtained here indicate that the mutagenicity of PAH is influenced not only by NER, but also by replication bypass fidelity. This has been postulated earlier based on results using in vitro enzyme assays, but is now also being recognized in terms of forward mutations in intact mammalian cells.
Citation:
DNA Repair (Amst.) 2008, 7 (8):1202-1212
Journal:
DNA repair
Issue Date:
2-Aug-2008
URI:
http://hdl.handle.net/10146/77381
DOI:
10.1016/j.dnarep.2008.03.022
PubMed ID:
18479980
Additional Links:
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6X17-4SH1J0G-1&_user=1843694&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=d549f87361b5a7a56939d2ecfff06b30
Type:
Article
Language:
en
ISSN:
1568-7864
Sponsors:
We would like to thank Dr. L. Thompson (LLNL, Livermore, CA, USA) for the Chinese hamster ovary cell lines used in this study. The XEM2 cells were a generous gift from Prof. Dr. J. Döhmer, GenPharmTox BioTech AG, 82152 Planegg/Martinsried (Germany). We would also like to show our gratitude towards Dr. M. Hunt for valuable improvements of the manuscript. This work was supported by the European Commission, projects; AMBIPAH QLK4-CT-2002-02402, DIEPHY FOOD-CT-2003-505609 and ECNIS (FOOD-CT-2005-513943).
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorLagerqvist, Anne-
dc.contributor.authorHakansson, Daniel-
dc.contributor.authorProchazka, Gabriela-
dc.contributor.authorLundin, Cecilia-
dc.contributor.authorDreij, Kristian-
dc.contributor.authorSegerback, Dan-
dc.contributor.authorJernstrom, Bengt-
dc.contributor.authorTornqvist, Margareta-
dc.contributor.authorSeidel, Albrecht-
dc.contributor.authorErixon, Klaus-
dc.contributor.authorJenssen, Dag-
dc.date.accessioned2009-08-14T11:24:42Z-
dc.date.available2009-08-14T11:24:42Z-
dc.date.issued2008-08-02-
dc.identifier.citationDNA Repair (Amst.) 2008, 7 (8):1202-1212en
dc.identifier.issn1568-7864-
dc.identifier.pmid18479980-
dc.identifier.doi10.1016/j.dnarep.2008.03.022-
dc.identifier.urihttp://hdl.handle.net/10146/77381-
dc.description.abstractMutations induced by polycyclic aromatic hydrocarbons (PAH) are expected to be produced when error-prone DNA replication occurs across unrepaired DNA lesions formed by reactive PAH metabolites such as diol epoxides. The mutagenicity of the two PAH-diol epoxides (+)-anti-7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) and (+/-)-anti-11,12-dihydroxy-13,14-epoxy-11,12,13,14-tetrahydrodibenzo[a,l]pyrene (DBPDE) was compared in nucleotide excision repair (NER) proficient and deficient hamster cell lines. We applied the (32)P-postlabelling assay to analyze adduct levels and the hprt gene mutation assay for monitoring mutations. It was found that the mutagenicity per target dose was 4 times higher for DBPDE compared to BPDE in NER proficient cells while in NER deficient cells, the mutagenicity per target dose was 1.4 times higher for BPDE. In order to investigate to what extent the mutagenicity of the different adducts in NER proficient cells was influenced by repair or replication bypass, we measured the overall NER incision rate, the rate of adduct removal, the rate of replication bypass and the frequency of induced recombination in the hprt gene. The results suggest that NER of BPDE lesions are 5 times more efficient than for DBPDE lesions, in NER proficient cells. However, DBPDE adducts block replication more efficiently and also induce 6 times more recombination events in the hprt gene than adducts of BPDE, suggesting that DBPDE adducts are, to a larger extent, bypassed by homologous recombination. The results obtained here indicate that the mutagenicity of PAH is influenced not only by NER, but also by replication bypass fidelity. This has been postulated earlier based on results using in vitro enzyme assays, but is now also being recognized in terms of forward mutations in intact mammalian cells.en
dc.description.sponsorshipWe would like to thank Dr. L. Thompson (LLNL, Livermore, CA, USA) for the Chinese hamster ovary cell lines used in this study. The XEM2 cells were a generous gift from Prof. Dr. J. Döhmer, GenPharmTox BioTech AG, 82152 Planegg/Martinsried (Germany). We would also like to show our gratitude towards Dr. M. Hunt for valuable improvements of the manuscript. This work was supported by the European Commission, projects; AMBIPAH QLK4-CT-2002-02402, DIEPHY FOOD-CT-2003-505609 and ECNIS (FOOD-CT-2005-513943).en
dc.language.isoenen
dc.relation.urlhttp://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6X17-4SH1J0G-1&_user=1843694&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=d549f87361b5a7a56939d2ecfff06b30en
dc.subjectBenzo[a]pyreneen
dc.subjectDibenzo[a,l]pyreneen
dc.subjectNucleotide excision repairen
dc.subjectDNA adductsen
dc.subjectReplication bypassen
dc.subjectMutationsen
dc.subject.mesh7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide-
dc.subject.meshAnimals-
dc.subject.meshBenzopyrenes-
dc.subject.meshCell Line-
dc.subject.meshChromatography, Thin Layer-
dc.subject.meshCricetinae-
dc.subject.meshCricetulus-
dc.subject.meshDNA Repair-
dc.subject.meshDNA Replication-
dc.subject.meshDose-Response Relationship, Drug-
dc.subject.meshEpoxy Compounds-
dc.subject.meshHalf-Life-
dc.subject.meshMutation-
dc.titleBoth replication bypass fidelity and repair efficiency influence the yield of mutations per target dose in intact mammalian cells induced by benzo[a]pyrene-diol-epoxide and dibenzo[a,l]pyrene-diol-epoxide.en
dc.typeArticleen
dc.identifier.journalDNA repairen
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