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FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing.

ABSTRACT: Genome replication mapping methods profile cell populations, masking cell-to-cell heterogeneity. Here, we describe FORK-seq, a nanopore sequencing method to map replication of single DNA molecules at 200-nucleotide resolution. By quantifying BrdU incorporation along pulse-chased replication intermediates from Saccharomyces cerevisiae, we orient 58,651 replication tracks reproducing population-based replication directionality profiles and map 4964 and 4485 individual initiation and termination events, respectively. Although most events cluster at known origins and fork merging zones, 9% and 18% of initiation and termination events, respectively, occur at many locations previously missed. Thus, FORK-seq reveals the full extent of cell-to-cell heterogeneity in DNA replication.

SUBMITTER: Hennion M

PROVIDER: S-EPMC7251829 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing.

Hennion Magali M Arbona Jean-Michel JM Lacroix Laurent L Cruaud Corinne C Theulot Bertrand B Tallec Benoît Le BL Proux Florence F Wu Xia X Novikova Elizaveta E Engelen Stefan S Lemainque Arnaud A Audit Benjamin B Hyrien Olivier O

Genome biology 20200526 1

Genome replication mapping methods profile cell populations, masking cell-to-cell heterogeneity. Here, we describe FORK-seq, a nanopore sequencing method to map replication of single DNA molecules at 200-nucleotide resolution. By quantifying BrdU incorporation along pulse-chased replication intermediates from Saccharomyces cerevisiae, we orient 58,651 replication tracks reproducing population-based replication directionality profiles and map 4964 and 4485 individual initiation and termination ev ...[more]

PMID: 32456659

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