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. Author manuscript; available in PMC: 2009 May 18.
Published in final edited form as: J Biol Chem. 2008 Dec 30;284(9):5654–5661. doi: 10.1074/jbc.M806973200

Fig. 5. Strand displacement activity of RRAA mutant is substantially reduced compared to WT.

Fig. 5

Autoradiographs showing strand displacement of a labelled 24mer annealed to a 50mer to produce a template with a 3′ ssDNA region. Lanes are: (“boil”), substrate boiled for 5 mins in the absence of protein; (“no protein”), substrate alone; (“ice”), reaction incubated on ice; (“- ATP’), reaction without ATP; (“+EDTA”), reaction in the presence of 10 mM EDTA. Remaining lanes are reactions containing increasing concentrations of MthMCM protein indicated in nM (monomer). (A) WT, (B) RRAA, (C) R137A. (D) Quantification of DNA unwinding activity from experiments shown in (A) – (C). Unwinding was calculated as a percentage of 24mer displaced from the 50mer template. All points are an average of at least three experiments and error bars show the standard deviation from the mean. WT, closed circles, continuous line; R137A, open diamond, continuous line; RRAA, open circles, broken line.