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We performed a comparative analysis of replication origin activation by genome-wide single-stranded DNA mapping in two yeast strains challenged by hydroxyurea, an inhibitor of the ribonucleotide reductase. We gained understanding of the impact on origin activation by three factors: S-phase checkpoint control, DNA sequence polymorphisms, and relative positioning of origin and transcription unit. Wild type W303 showed a significant reduction of fork progression accompanied by an elevated level ofdoi:10.1371/journal.pone.0263569 pmid:35157703 pmcid:PMC8843211 fatcat:vyaevww4lzbdlpd4qsxoxx5msu