Genome-wide mapping of individual replication fork velocities using nanopore sequencing

Bertrand Theulot, Laurent Lacroix (), Jean-Michel Arbona, Gael A. Millot, Etienne Jean, Corinne Cruaud, Jade Pellet, Florence Proux, Magali Hennion, Stefan Engelen, Arnaud Lemainque, Benjamin Audit, Olivier Hyrien () and Benoît Tallec ()
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Bertrand Theulot: Université PSL
Laurent Lacroix: Université PSL
Jean-Michel Arbona: Université Claude Bernard Lyon 1
Gael A. Millot: Université Paris Cité, Bioinformatics and Biostatistics Hub
Etienne Jean: Université PSL
Corinne Cruaud: Université Paris-Saclay
Jade Pellet: Université PSL
Florence Proux: Université PSL
Magali Hennion: Université Paris Cité, Epigenetics and Cell Fate, UMR7216, CNRS
Stefan Engelen: Univ. Evry, Université Paris-Saclay
Arnaud Lemainque: Université Paris-Saclay
Benjamin Audit: ENSL, CNRS, Laboratoire de physique
Olivier Hyrien: Université PSL
Benoît Tallec: Université PSL

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Little is known about replication fork velocity variations along eukaryotic genomes, since reference techniques to determine fork speed either provide no sequence information or suffer from low throughput. Here we present NanoForkSpeed, a nanopore sequencing-based method to map and extract the velocity of individual forks detected as tracks of the thymidine analogue bromodeoxyuridine incorporated during a brief pulse-labelling of asynchronously growing cells. NanoForkSpeed retrieves previous Saccharomyces cerevisiae mean fork speed estimates (≈2 kb/min) in the BT1 strain exhibiting highly efficient bromodeoxyuridine incorporation and wild-type growth, and precisely quantifies speed changes in cells with altered replisome progression or exposed to hydroxyurea. The positioning of >125,000 fork velocities provides a genome-wide map of fork progression based on individual fork rates, showing a uniform fork speed across yeast chromosomes except for a marked slowdown at known pausing sites.

Date: 2022
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DOI: 10.1038/s41467-022-31012-0

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