Fig. 7: RPA-SSL and SKYD mutants are impaired in DNA end resection.
From: Distinct RPA domains promote recruitment and the helicase-nuclease activities of Dna2
a A representative experiment to monitor DNA end resection by Sgs1, Dna2, and increasing concentrations of yeast RPA variants using 2.2-knt-long dsDNA (0.5 nM, in molecules). The reaction buffer contained 100 mM NaCl. Red asterisks indicate random radioactive labels on the DNA. b Quantification of overall substrate utilization from experiments such as shown in panel a. Error bars, SEM; n = 3. c A model showing that RPA first recruits Dna2 to 5′-overhanged DNA (1). Subsequently, RPA remains a component of the nucleoprotein complex and stimulates both nuclease (Nuc) and ATPase-driven translocase (Hel) activities of Dna2 (2). d A model depicting domains of the RPA large subunit, Rfa1, and their involvement in ssDNA binding, recruitment of Dna2 and stimulation of nuclease and helicase activities of Dna2, respectively.
