Genomic patterns of transcription-replication interactions in mouse primary B cells

Commodore P. St Germain, Hongchang Zhao, Vrishti Sinha, Lionel A. Sanz, Frédéric Chédin, Jacqueline H. Barlow

Research output: Contribution to journalArticlepeer-review

Abstract

Conflicts between transcription and replication machinery are a potent source of replication stress and genome instability; however, no technique currently exists to identify endogenous genomic locations prone to transcription-replication interactions. Here, we report a novel method to identify genomic loci prone to transcription-replication interactions termed transcription-replication immunoprecipitation on nascent DNA sequencing, TRIPn-Seq. TRIPn-Seq employs the sequential immunoprecipitation of RNA polymerase 2 phosphorylated at serine 5 (RNAP2s5) followed by enrichment of nascent DNA previously labeled with bromodeoxyuridine. Using TRIPn-Seq, we mapped 1009 unique transcription-replication interactions (TRIs) in mouse primary B cells characterized by a bimodal pattern of RNAP2s5, bidirectional transcription, an enrichment of RNA:DNA hybrids, and a high probability of forming G-quadruplexes. TRIs are highly enriched at transcription start sites and map to early replicating regions. TRIs exhibit enhanced Replication Protein A association and TRI-associated genes exhibit higher replication fork termination than control transcription start sites, two marks of replication stress. TRIs colocalize with double-strand DNA breaks, are enriched for deletions, and accumulate mutations in tumors. We propose that replication stress at TRIs induces mutations potentially contributing to age-related disease, as well as tumor formation and development.

Original languageEnglish (US)
Pages (from-to)2051-2073
Number of pages23
JournalNucleic acids research
Volume50
Issue number4
DOIs
StatePublished - Feb 28 2022
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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