Extended Data Fig. 7: SAMHD1 binds MRE11 and stimulates its nuclease activity.
From: SAMHD1 acts at stalled replication forks to prevent interferon induction
a, Various fluorescently labelled DNA substrates (ssDNA, dsDNA, dsDNA with a 5′ overhang, forked DNA and reversed fork DNA of the nucleotide lengths indicated) were incubated with a range of concentrations of purified SAMHD1 and the formation of protein–DNA complexes was determined by electrophoretic mobility shift assay (EMSA) as described in the Supplementary Information. Representative gel shift images are shown (n = 3). b, Quantification of the EMSA in a. c, SAMHD1 stimulates the nuclease activity of yeast MRE11, but not that of bacterial ExoIII and human FEN1 and DNA2. Error bars denote s.d. of three independent experiments. d, e, SAMHD1 binds MRE11 and RPA as monitored by microscale thermophoresis assay. Error bars denote s.d. of three independent experiments. f, Co-immunoprecipitation of Samhd1 and RPA from Xenopus egg extracts. Western blots are shown of the input egg extract (XE), the proteins immunoprecipitated (IP) by a control antibody (IgG), and an antibody against Samhd1 (αSAM), and the proteins that remained unbound (flow through, FT). A representative experiment is shown (n = 2). g, Co-immunoprecipitation of Samhd1 and CtIP from Xenopus egg extracts, as in f and described in the Supplementary Information. A representative experiment is shown (n = 2).
