Nucleosome-directed replication origin licensing independent of a consensus DNA sequence

Sai Li, Michael R. Wasserman, Olga Yurieva, Lu Bai, Michael E. O’Donnell () and Shixin Liu ()
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Sai Li: The Rockefeller University
Michael R. Wasserman: The Rockefeller University
Olga Yurieva: The Rockefeller University
Lu Bai: The Pennsylvania State University
Michael E. O’Donnell: The Rockefeller University
Shixin Liu: The Rockefeller University

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract The numerous enzymes and cofactors involved in eukaryotic DNA replication are conserved from yeast to human, and the budding yeast Saccharomyces cerevisiae (S.c.) has been a useful model organism for these studies. However, there is a gap in our knowledge of why replication origins in higher eukaryotes do not use a consensus DNA sequence as found in S.c. Using in vitro reconstitution and single-molecule visualization, we show here that S.c. origin recognition complex (ORC) stably binds nucleosomes and that ORC-nucleosome complexes have the intrinsic ability to load the replicative helicase MCM double hexamers onto adjacent nucleosome-free DNA regardless of sequence. Furthermore, we find that Xenopus laevis nucleosomes can substitute for yeast ones in engaging with ORC. Combined with re-analyses of genome-wide ORC binding data, our results lead us to propose that the yeast origin recognition machinery contains the cryptic capacity to bind nucleosomes near a nucleosome-free region and license origins, and that this nucleosome-directed origin licensing paradigm generalizes to all eukaryotes.

Date: 2022
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DOI: 10.1038/s41467-022-32657-7

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