Cooperative control of holliday junction resolution and DNA repair by the SLX1 and MUS81-EME1 nucleases

Mol Cell. 2013 Oct 24;52(2):221-33. doi: 10.1016/j.molcel.2013.08.036. Epub 2013 Sep 26.

Abstract

Holliday junctions (HJs) are X-shaped DNA structures that arise during homologous recombination, which must be removed to enable chromosome segregation. The SLX1 and MUS81-EME1 nucleases can both process HJs in vitro, and they bind in close proximity on the SLX4 scaffold, hinting at possible cooperation. However, the cellular roles of mammalian SLX1 are not yet known. Here, we use mouse genetics and structure function analysis to investigate SLX1 function. Disrupting the murine Slx1 and Slx4 genes revealed that they are essential for HJ resolution in mitotic cells. Moreover, SLX1 and MUS81-EME1 act together to resolve HJs in a manner that requires tethering to SLX4. We also show that SLX1, like MUS81-EME1, is required for repair of DNA interstrand crosslinks, but this role appears to be independent of HJ cleavage, at least in mouse cells. These findings shed light on HJ resolution in mammals and on maintenance of genome stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Cells, Cultured
  • DNA / genetics
  • DNA / metabolism
  • DNA Repair*
  • DNA, Cruciform*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Embryo, Mammalian / cytology
  • Endodeoxyribonucleases / genetics
  • Endodeoxyribonucleases / metabolism*
  • Endonucleases / genetics
  • Endonucleases / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • HEK293 Cells
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Models, Genetic
  • Molecular Sequence Data
  • Protein Binding
  • RNA Interference
  • Recombinases / genetics
  • Recombinases / metabolism
  • Sequence Homology, Amino Acid

Substances

  • DNA, Cruciform
  • DNA-Binding Proteins
  • Recombinases
  • DNA
  • Eme1 protein, mouse
  • Endodeoxyribonucleases
  • Endonucleases
  • Mus81 protein, mouse
  • SLX1 protein, mouse
  • Slx4 protein, mouse