Crystal structure and interactions of the Tof1-Csm3 (Timeless-Tipin) fork protection complex

Nucleic Acids Res. 2020 Jul 9;48(12):6996-7004. doi: 10.1093/nar/gkaa456.

Abstract

The Tof1-Csm3 fork protection complex has a central role in the replisome-it promotes the progression of DNA replication forks and protects them when they stall, while also enabling cohesion establishment and checkpoint responses. Here, I present the crystal structure of the Tof1-Csm3 complex from Chaetomium thermophilum at 3.1 Å resolution. The structure reveals that both proteins together form an extended alpha helical repeat structure, which suggests a mechanical or scaffolding role for the complex. Expanding on this idea, I characterize a DNA interacting region and a cancer-associated Mrc1 binding site. This study provides the molecular basis for understanding the functions of the Tof1-Csm3 complex, its human orthologue the Timeless-Tipin complex and additionally the Drosophila circadian rhythm protein Timeless.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / ultrastructure*
  • Chaetomium / ultrastructure
  • Crystallography, X-Ray
  • DNA Replication / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / ultrastructure*
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / ultrastructure*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / ultrastructure

Substances

  • Cell Cycle Proteins
  • Csm3p protein, S cerevisiae
  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Multiprotein Complexes
  • Saccharomyces cerevisiae Proteins
  • TIMELESS protein, human
  • TOF1 protein, S cerevisiae
  • Tipin protein, human