Skip to main content
Figure 7 | BMC Molecular Biology

Figure 7

From: Length-dependent degradation of single-stranded 3' ends by the Werner syndrome protein (WRN): implications for spatial orientation and coordinated 3' to 5' movement of its ATPase/helicase and exonuclease domains

Figure 7

DNA binding and exonuclease activities of the isolated exonuclease domain of WRN. A) Wild type WRN or the exonuclease domain-only deletion mutant WRNΔ369-1432 (0.9, 3.6, 7.2, 15 or 30 nM each) was incubated for 5 min at 37°C with G45 (0.025 nM) in the presence of ATPγS (1 mM). Unbound DNA and DNA-protein complexes were separated using EMSA as described in Methods. B) Wild type WRN or WRNΔ369-1432 (3, 6 or 18 nM each) was incubated for 15 min at 37°C with single-stranded, radiolabeled G35 (0.1 nM, left) or G40 (0.1 nM, right) in the absence of ATP. C) Similarly, WRNΔ369-1432 (120 nM) was also incubated with either G35 or G40 (0.1 nM each) in the absence or presence of ATP or ATPγS (1 mM) for 15 min at 37°C. D) Standard exonuclease assays were performed on 10 nt 3' overhang substrate (labeled C55/G45, 0.5 nM) with WRNΔ369-1432 (20, 40, 80 or 120 nM) in 1 mM ATP for 10 min at 37°C (left). Similar reactions were performed with 10 nt overhang substrate (0.25 nM) and WRNΔ369-1432 (120 nM) in the absence or presence of ATP or ATPγS (1 mM) for10 min at 37°C (right). For B-D, exonuclease assays were performed as described in Methods. Lengths of the original substrates (in nt) are indicated at left of each panel.

Back to article page