Fig. 3: Nse1-3-4 subcomplex contributes to Smc5/6’s association with DNA.
From: Smc5/6’s multifaceted DNA binding capacities stabilize branched DNA structures
a Schematic of the Smc5/6 5-mer complex, which lacks the Nse1-3-4 subcomplex. b Representative kymograph of a λ DNA tether being stretched from LF to HF in the presence of 20 nM Alexa555-Smc5/6 5-mer (green) without ATP. dsDNA (blue), ssDNA (red), and junction DNA (green) regions of the tether are indicated to the right of the kymograph. c Fluorescence signals of Smc5/6 5-mer and 8-mer complexes on dsDNA under LF without or with ATP (no RPA). Bar heights indicate the group mean and error bars represent standard deviation. P values were determined from two-tailed unpaired t-tests with Welch’s correction (**P < 0.01; ***P < 0.001). P = 0.0002 for −ATP condition, and P = 0.0028 for +ATP condition. Sample sizes are: 5-mer dsDNALF −ATP (n = 6); 8-mer dsDNALF −ATP (n = 6); 5-mer dsDNALF + ATP (n = 5); and 8-mer dsDNALF + ATP (n = 6), where n indicates the number of regions analyzed. d Fluorescence signals of Smc5/6 5-mer and 8-mer complexes on ssDNA under HF in the presence of ATP without or with RPA. Bar heights indicate the group mean and error bars represent standard deviation. P values were determined from two-tailed unpaired t-tests with Welch’s correction (****P < 0.0001). Sample sizes are: 5-mer ssDNAHF −RPA (n = 11); 5-mer ssDNAHF + RPA (n = 6); 8-mer ssDNAHF −RPA (n = 12); 8-mer ssDNAHF + RPA (n = 5), where n indicates the number of regions analyzed. e Fluorescence signals of Smc5/6 5-mer and 8-mer complexes on junction DNA under HF in the presence of ATP without or with RPA. Bar heights indicate the group mean and error bars represent standard deviation. P values were determined from two-tailed unpaired t-tests with Welch’s correction (****P < 0.0001). Sample sizes are: 5-mer JunctionHF −RPA (n = 16); 5-mer JunctionHF + RPA (n = 12); 8-mer JunctionHF −RPA (n = 21); and 8-mer JunctionHF + RPA (n = 13), where n indicates the number of regions analyzed. Source data for panels c, d, and e are provided within the Source Data file.
