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. 2001 Mar 15;29(6):1317–1325. doi: 10.1093/nar/29.6.1317

Figure 3.

Figure 3

Characterization of hMre11 DNA-binding properties. (A) Radiolabeled 50 nt ssDNA (MJ19; left) or dsDNA (MJ19/MJ20; right) was incubated at 1.25 nM with hMre11 at 50 nM. After 15 min incubation, the indicated nucleotide excess non-radioactively-labeled competitor DNA was added and incubation was continued for an additional 15 min. Competitor DNA was single-stranded oligonucleotide (MJ19) or circular φX174 ssDNA (left) and blunt-ended double-stranded oligonucleotide (MJ19/MJ20) or circular φX174 dsDNA (right). Lane ‘–’, incubation without hMre11. Samples were analyzed by native PAGE, followed by autoradiography. (B) Radiolabeled 50 nt ssDNA (MJ19) was incubated at 1.25 nM with 50 nM hMre11 as in (A). Non-labeled, non-identical ssDNA (DG73) or dsDNA (DG73/DG74) oligonucleotide DNA was used as a competitor. Values on the y-axis are reciprocal fractions of DNA–protein complex, normalized to the fraction of DNA–protein complex in non-competed reactions (Materials and Methods). Bars indicate standard deviations of mean values of triplicate reactions. Slopes of linear regression lines through the datapoints are indicated.