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. 2014 Sep 8;42(17):11083–11091. doi: 10.1093/nar/gku803

Figure 3.

Figure 3.

Stimulation of BLM helicase processivity by the TR complex and DNA2-D277A. (A) BLM-mediated DNA unwinding was examined with 5 nM BLM and the addition of DNA2-D277A or DNA2-D277A/K654E (5 or 20 nM). Reactions were incubated for 10 min at 37°C (B) Binding of an 80-mer dsDNA by BLM (7.5, 10.5 or 15 nM) without or with DNA2-D277A (10 or 20 nM). The data shown are the average from three independent experiments and the error bars represent 1 SD. (C) A randomly radiolabeled 2 kb dsDNA (0.5 nM ends) was incubated with BLM (5.5 nM) and RPA (200 nM), and 10-fold excess unlabeled dsDNA (identical to the labeled substrate in sequence) was added at the 2-min point along with the TR complex (5 nM) or DNA2-D277A (16.5 nM). Samples were taken at 2.5 min intervals and analyzed. Quantification of the results is shown below. (D) Resection of a 2 kb dsDNA (0.5 nM ends) by BLM (2 nM), DNA2 (10 nM) and RPA (200 nM) was examined without or with the TR complex (5 nM). The average data from three independent experiments are shown and the error bars represent 1 SD.