Dynamic relocalization of replication origins by Fkh1 requires execution of DDK function and Cdc45 loading at origins

Elife. 2019 May 14;8:e45512. doi: 10.7554/eLife.45512.

Abstract

Chromosomal DNA elements are organized into spatial domains within the eukaryotic nucleus. Sites undergoing DNA replication, high-level transcription, and repair of double-strand breaks coalesce into foci, although the significance and mechanisms giving rise to these dynamic structures are poorly understood. In S. cerevisiae, replication origins occupy characteristic subnuclear localizations that anticipate their initiation timing during S phase. Here, we link localization of replication origins in G1 phase with Fkh1 activity, which is required for their early replication timing. Using a Fkh1-dependent origin relocalization assay, we determine that execution of Dbf4-dependent kinase function, including Cdc45 loading, results in dynamic relocalization of a replication origin from the nuclear periphery to the interior in G1 phase. Origin mobility increases substantially with Fkh1-driven relocalization. These findings provide novel molecular insight into the mechanisms that govern dynamics and spatial organization of DNA replication origins and possibly other functional DNA elements.

Keywords: S. cerevisiae; chromosome; chromosomes; forkhead; gene expression; nuclear mobility; nucleus; replication origin; replication timing.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Cycle Proteins / metabolism*
  • DNA Repair
  • DNA Replication
  • DNA-Binding Proteins / metabolism*
  • Forkhead Transcription Factors / metabolism*
  • Nuclear Proteins / metabolism*
  • Replication Origin*
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription, Genetic

Substances

  • CDC45 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Dbf4 protein, S cerevisiae
  • Fkh1 protein, S cerevisiae
  • Forkhead Transcription Factors
  • Nuclear Proteins
  • Saccharomyces cerevisiae Proteins

Associated data

  • Dryad/10.5061/dryad.7bm444s