title: A recombinase paralog from the hyperthermophilic crenarchaeon Sulfolobus solfataricus enhances SsoRadA ssDNA binding and strand displacement.
abstract: Homologous recombination (HR) is a major pathway for the repair of double-strand DNA breaks, a highly deleterious form of DNA damage. The main catalytic protein in HR is the essential RecA-family recombinase, which is conserved across all three domains of life. Eukaryotes and archaea encode varying numbers of proteins paralogous to their main recombinase. Although there is increasing evidence for the functions of some of these paralog proteins, overall their mechanism of action remains largely unclear. Here we present the first biochemical characterization of one of the paralog proteins, SsoRal3, from the crenarchaeaon Sulfolobus solfataricus. The SsoRal3 protein is a ssDNA-dependent ATPase that can catalyze strand invasion at both saturating and subsaturating concentrations. It can bind both ssDNA and dsDNA, but its binding preference is altered by the presence or absence of ATP. Addition of SsoRal3 to SsoRadA nucleoprotein filaments reduces total ATPase activity. Subsaturating concentrations of SsoRal3 increase the ssDNA binding activity of SsoRadA approximately 9-fold and also increase the persistence of SsoRadA catalyzed strand invasion products. Overall, these results suggest that SsoRal3 functions to stabilize the SsoRadA presynaptic filament.
authors.affiliation: School of Molecular Biosciences, Washington State University, Pullman, WA 99163, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: Graham
authors.forename: Cynthia A
authors.initials: CA
authors.lastname: Haseltine
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics* / Q000378:metabolism*; D004789:Enzyme Activation; D015870:Gene Expression; D059765:Homologous Recombination; D011485:Protein Binding; D011938:Rec A Recombinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D048229:Sulfolobus solfataricus / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D011938:Rec A Recombinases; D000251:Adenosine Triphosphatases