A role for SSRP1 in recombination-mediated DNA damage response

J Cell Biochem. 2009 Oct 1;108(2):508-18. doi: 10.1002/jcb.22280.


A possible role for structure-specific recognition protein 1 (SSRP1) in replication-associated repair processes has previously been suggested based on its interaction with several DNA repair factors and the replication defects observed in SSRP1 mutants. In this study, we investigated the potential role of SSRP1 in association with DNA repair mediated by homologous recombination (HR), one of the pathways involved in repairing replication-associated DNA damage, in mammalian cells. Surprisingly, over-expression of SSRP1 reduced the number of hprt(+) recombinants generated via HR both spontaneously and upon hydroxyurea (HU) treatment, whereas knockdown of SSRP1 resulted in an increase of HR events in response to DNA double-strand break formation. In correlation, we found that the depletion of SSRP1 in HU-treated human cells elevated the number of Rad51 and H2AX foci, while over-expression of the wild-type SSRP1 markedly reduced HU-induced Rad51 foci formation. We also found that SSRP1 physically interacts with a key HR repair protein, Rad54 both in vitro and in vivo. Further, branch migration studies demonstrated that SSRP1 inhibits Rad54-promoted branch migration of Holliday junctions in vitro. Taken together, our data suggest a functional role for SSRP1 in spontaneous and replication-associated DNA damage response by suppressing avoidable HR repair events.

Publication types

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

MeSH terms

  • Animals
  • Blotting, Western
  • Cricetinae
  • Cricetulus
  • DNA Breaks, Double-Stranded
  • DNA Damage
  • DNA Helicases
  • DNA Repair*
  • DNA, Cruciform / metabolism
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / deficiency
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Gene Knockdown Techniques
  • Genes, Reporter
  • High Mobility Group Proteins / biosynthesis
  • High Mobility Group Proteins / deficiency
  • High Mobility Group Proteins / genetics
  • High Mobility Group Proteins / physiology*
  • Histones / metabolism
  • Humans
  • Hydroxyurea
  • Mutant Proteins / isolation & purification
  • Mutant Proteins / metabolism
  • Nuclear Proteins / metabolism
  • Peptide Fragments
  • Plasmids
  • Protein Binding
  • RNA, Small Interfering
  • Rad51 Recombinase / metabolism
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic*
  • Transcriptional Elongation Factors / biosynthesis
  • Transcriptional Elongation Factors / deficiency
  • Transcriptional Elongation Factors / genetics
  • Transcriptional Elongation Factors / physiology*
  • Transfection


  • DNA, Cruciform
  • DNA-Binding Proteins
  • H2AX protein, human
  • High Mobility Group Proteins
  • Histones
  • Mutant Proteins
  • Nuclear Proteins
  • Peptide Fragments
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • SSRP1 protein, human
  • Transcriptional Elongation Factors
  • Rad51 Recombinase
  • DNA Helicases
  • RAD54L protein, human
  • Hydroxyurea