CDK activity provides temporal and quantitative cues for organizing genome duplication

PLoS Genet. 2018 Feb 21;14(2):e1007214. doi: 10.1371/journal.pgen.1007214. eCollection 2018 Feb.

Abstract

In eukaryotes, the spatial and temporal organization of genome duplication gives rise to distinctive profiles of replication origin usage along the chromosomes. While it has become increasingly clear that these programs are important for cellular physiology, the mechanisms by which they are determined and modulated remain elusive. Replication initiation requires the function of cyclin-dependent kinases (CDKs), which associate with various cyclin partners to drive cell proliferation. Surprisingly, although we possess detailed knowledge of the CDK regulators and targets that are crucial for origin activation, little is known about whether CDKs play a critical role in establishing the genome-wide pattern of origin selection. We have addressed this question in the fission yeast, taking advantage of a simplified cell cycle network in which cell proliferation is driven by a single cyclin-CDK module. This system allows us to precisely control CDK activity in vivo using chemical genetics. First, in contrast to previous reports, our results clearly show that distinct cyclin-CDK pairs are not essential for regulating specific subsets of origins and for establishing a normal replication program. Importantly, we then demonstrate that the timing at which CDK activity reaches the S phase threshold is critical for the organization of replication in distinct efficiency domains, while the level of CDK activity at the onset of S phase is a dose-dependent modulator of overall origin efficiencies. Our study therefore implicates these different aspects of CDK regulation as versatile mechanisms for shaping the architecture of DNA replication across the genome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle / genetics
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Cell Division / genetics*
  • Cyclin-Dependent Kinases / physiology*
  • DNA Replication*
  • Genome, Fungal / genetics*
  • Organisms, Genetically Modified
  • Phosphorylation
  • Replication Origin / genetics
  • S Phase / genetics
  • Schizosaccharomyces / cytology
  • Schizosaccharomyces / enzymology
  • Schizosaccharomyces / genetics
  • Signal Transduction / genetics

Substances

  • Cell Cycle Proteins
  • Cyclin-Dependent Kinases

Grants and funding

This study was supported by grants to PYJW from the Centre National pour la Recherche Scientific (CNRS, www.cnrs.fr) / Institut national de la santé et de la recherche médicale (INSERM, www.inserm.fr) through the ATIP-Avenir program and the Fondation pour la Recherche Médicale (FRM, www.frm.org) through the “Amorçage de jeunes équipes” program. AP was supported by funding from the Région Bretagne (www.bretagne.bzh), the Fondation ARC (www.fondation-arc.org), and the Institute of Genetics and Development of Rennes (igdr.univ-rennes1.fr). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.