TY - JOUR
T1 - Genome-wide mapping of 8-oxo-7,8-dihydro-2′-deoxyguanosine reveals accumulation of oxidatively-generated damage at DNA replication origins within transcribed long genes of mammalian cells
AU - Amente, Stefano
AU - Di Palo, Giacomo
AU - Scala, Giovanni
AU - Castrignanò, Tiziana
AU - Gorini, Francesca
AU - Cocozza, Sergio
AU - Moresano, Angela
AU - Pucci, Piero
AU - Ma, Bin
AU - Stepanov, Irina
AU - Lania, Luigi
AU - Pelicci, Pier Giuseppe
AU - Dellino, Gaetano Ivan
AU - Majello, Barbara
N1 - Publisher Copyright:
© 2018 The Author(s).
PY - 2019/1/10
Y1 - 2019/1/10
N2 - 8-Oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is one of the major DNA modifications and a potent pre-mutagenic lesion prone to mispair with 2′-deoxyadenosine (dA). Several thousand residues of 8-oxodG are constitutively generated in the genome of mammalian cells, but their genomic distribution has not yet been fully characterized. Here, by using OxiDIP-Seq, a highly sensitive methodology that uses immuno-precipitation with efficient anti-8-oxodG antibodies combined with high-throughput sequencing, we report the genome-wide distribution of 8-oxodG in human non-tumorigenic epithelial breast cells (MCF10A), and mouse embryonic fibroblasts (MEFs). OxiDIP-Seq revealed sites of 8-oxodG accumulation overlapping with Î 3H2AX ChIP-Seq signals within the gene body of transcribed long genes, particularly at the DNA replication origins contained therein. We propose that the presence of persistent single-stranded DNA, as a consequence of transcription-replication clashes at these sites, determines local vulnerability to DNA oxidation and/or its slow repair. This oxidatively-generated damage, likely in combination with other kinds of lesion, might contribute to the formation of DNA double strand breaks and activation of DNA damage response.
AB - 8-Oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is one of the major DNA modifications and a potent pre-mutagenic lesion prone to mispair with 2′-deoxyadenosine (dA). Several thousand residues of 8-oxodG are constitutively generated in the genome of mammalian cells, but their genomic distribution has not yet been fully characterized. Here, by using OxiDIP-Seq, a highly sensitive methodology that uses immuno-precipitation with efficient anti-8-oxodG antibodies combined with high-throughput sequencing, we report the genome-wide distribution of 8-oxodG in human non-tumorigenic epithelial breast cells (MCF10A), and mouse embryonic fibroblasts (MEFs). OxiDIP-Seq revealed sites of 8-oxodG accumulation overlapping with Î 3H2AX ChIP-Seq signals within the gene body of transcribed long genes, particularly at the DNA replication origins contained therein. We propose that the presence of persistent single-stranded DNA, as a consequence of transcription-replication clashes at these sites, determines local vulnerability to DNA oxidation and/or its slow repair. This oxidatively-generated damage, likely in combination with other kinds of lesion, might contribute to the formation of DNA double strand breaks and activation of DNA damage response.
KW - 8-Hydroxy-2'-Deoxyguanosine
KW - Animals
KW - Cell Line, Tumor
KW - Chromosome Mapping
KW - DNA/chemistry
KW - DNA Damage/genetics
KW - DNA Replication/genetics
KW - DNA, Single-Stranded/genetics
KW - Deoxyadenosines/genetics
KW - Deoxyguanosine/analogs & derivatives
KW - Fibroblasts/metabolism
KW - Genome/genetics
KW - Histones/genetics
KW - Humans
KW - Mice
KW - Oxidation-Reduction
KW - Replication Origin/genetics
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U2 - 10.1093/nar/gky1152
DO - 10.1093/nar/gky1152
M3 - Article
C2 - 30462294
AN - SCOPUS:85059795855
SN - 0305-1048
VL - 47
SP - 221
EP - 236
JO - Nucleic acids research
JF - Nucleic acids research
IS - 1
ER -