title: Conformational changes modulate the activity of human RAD51 protein.
abstract: Homologous recombination provides a major pathway for the repair of DNA double-strand breaks in mammalian cells. Defects in homologous recombination can lead to high levels of chromosomal translocations or deletions, which may promote cell transformation and cancer development. A key component of this process is RAD51. In comparison to RecA, the bacterial homologue, human RAD51 protein exhibits low-level strand-exchange activity in vitro. This activity can, however, be stimulated by the presence of high salt. Here, we have investigated the mechanistic basis for this stimulation. We show that high ionic strength favours the co-aggregation of RAD51-single-stranded DNA (ssDNA) nucleoprotein filaments with naked duplex DNA, to form a complex in which the search for homologous sequences takes place. High ionic strength allows differential binding of RAD51 to ssDNA and double-stranded DNA (dsDNA), such that ssDNA-RAD51 interactions are unaffected, whereas those between RAD51 and dsDNA are destabilized. Most importantly, high salt induces a conformational change in RAD51, leading to the formation of extended nucleoprotein filaments on ssDNA. These extended filaments mimic the active form of the Escherichia coli RecA-ssDNA filament that exhibits efficient strand-exchange activity.
authors.affiliation: Cancer Research UK, London Research Institute, Clare Hall Laboratories, South Mimms, Hertfordshire EN6 3LD, UK.
authors.forename: Yilun
authors.initials: Y
authors.lastname: Liu
authors.forename: Alicja Z
authors.initials: AZ
authors.lastname: Stasiak
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Masson
authors.forename: Michael J
authors.initials: MJ
authors.lastname: McIlwraith
authors.forename: Andrzej
authors.initials: A
authors.lastname: Stasiak
authors.forename: Stephen C
authors.initials: SC
authors.lastname: West
mesh_terms: D004247:DNA / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D009994:Osmolar Concentration; D011485:Protein Binding; D011487:Protein Conformation; D051135:Rad51 Recombinase
chemical_list: D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D004247:DNA; C495153:RAD51 protein, human; D051135:Rad51 Recombinase