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Figure 2 | BMC Molecular Biology

Figure 2

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 2

WRN binding to single-stranded DNA is length-dependent. For only these EMSA experiments, oligomers contained a 3'-PO4 group to block the exonuclease activity of wild type WRN. A) Wild type WRN (0.6, 1.2, 2.4, or 4.8 nM) was incubated with G80 (0.025 nM) in the presence or absence of ATP or ATPγS (1 mM), and free DNA and DNA-protein complexes were separated using EMSA as described in Methods. B) DNA-protein complexes formed by incubation of G80 (0.025 nM) with WRN (1.8 nM) in ATPγS were subsequently incubated with or without anti-WRN antibody as indicated and analyzed as in A. The slower mobility (supershift) of the protein-DNA complexes (denoted by asterisk) mediated by the anti-WRN antibody (lane 3) indicates that the complexes formed contain WRN. C) G80, G45, G40, G35, G30, or G24 (0.025 nM) was incubated without or with WRN (0.9, 1.8, or 3.6 nM) in ATPγS and products were analyzed as in A. D) From EMSA experiments as in C, quantitation and graphical representation of WRN binding to the G80 (), G45 (■), G40 (▲), G35 (), G30 (×), and G24 () oligomers. Each data point is the mean of three independent experiments, with the exception of G24 (two experiments).

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