Interaction of the Warsaw breakage syndrome DNA helicase DDX11 with the replication fork-protection factor Timeless promotes sister chromatid cohesion

PLoS Genet. 2018 Oct 10;14(10):e1007622. doi: 10.1371/journal.pgen.1007622. eCollection 2018 Oct.

Abstract

Establishment of sister chromatid cohesion is coupled to DNA replication, but the underlying molecular mechanisms are incompletely understood. DDX11 (also named ChlR1) is a super-family 2 Fe-S cluster-containing DNA helicase implicated in Warsaw breakage syndrome (WABS). Herein, we examined the role of DDX11 in cohesion establishment in human cells. We demonstrated that DDX11 interacts with Timeless, a component of the replication fork-protection complex, through a conserved peptide motif. The DDX11-Timeless interaction is critical for sister chromatid cohesion in interphase and mitosis. Immunofluorescence studies further revealed that cohesin association with chromatin requires DDX11. Finally, we demonstrated that DDX11 localises at nascent DNA by SIRF analysis. Moreover, we found that DDX11 promotes cohesin binding to the DNA replication forks in concert with Timeless and that recombinant purified cohesin interacts with DDX11 in vitro. Collectively, our results establish a critical role for the DDX11-Timeless interaction in coordinating DNA replication with sister chromatid cohesion, and have important implications for understanding the molecular basis of WABS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abnormalities, Multiple / genetics
  • Abnormalities, Multiple / metabolism
  • Abnormalities, Multiple / pathology
  • Cell Cycle Proteins / genetics*
  • Cell Cycle Proteins / metabolism
  • Chromatids / genetics*
  • Chromatids / metabolism
  • Chromosome Segregation / genetics
  • DEAD-box RNA Helicases / genetics*
  • DEAD-box RNA Helicases / metabolism
  • DNA / genetics
  • DNA / metabolism
  • DNA Helicases / genetics*
  • DNA Helicases / metabolism
  • DNA Replication / genetics*
  • Genomic Instability
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics*
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Protein Binding
  • Syndrome

Substances

  • Cell Cycle Proteins
  • Intracellular Signaling Peptides and Proteins
  • TIMELESS protein, human
  • DNA
  • DNA Helicases
  • DDX11 protein, human
  • DEAD-box RNA Helicases

Grant support

This work was supported by "Progetto Premiale-MIUR" TERABIO to [FMP]; “FaReBio di Qualità” fellowship from "Ministero della Economia" to [PP]; the Cancer Prevention and Research Institute of Texas [RP120717-P2 and RP160667-P2 to HY]; and the Welch Foundation [I-1441 to HY]. HY is an Investigator with the Howard Hughes Medical Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.