Movement of the RecG Motor Domain upon DNA Binding Is Required for Efficient Fork Reversal
AbstractRecG catalyzes reversal of stalled replication forks in response to replication stress in bacteria. The protein contains a fork recognition (“wedge”) domain that binds branched DNA and a superfamily II (SF2) ATPase motor that drives translocation on double-stranded (ds)DNA. The mechanism by which the wedge and motor domains collaborate to catalyze fork reversal in RecG and analogous eukaryotic fork remodelers is unknown. Here, we used electron paramagnetic resonance (EPR) spectroscopy to probe conformational changes between the wedge and ATPase domains in response to fork DNA binding by Thermotoga maritima RecG. Upon binding DNA, the ATPase-C lobe moves away from both the wedge and ATPase-N domains. This conformational change is consistent with a model of RecG fully engaged with a DNA fork substrate constructed from a crystal structure of RecG bound to a DNA junction together with recent cryo-electron microscopy (EM) structures of chromatin remodelers in complex with dsDNA. We show by mutational analysis that a conserved loop within the translocation in RecG (TRG) motif that was unstructured in the RecG crystal structure is essential for fork reversal and DNA-dependent conformational changes. Together, this work helps provide a more coherent model of fork binding and remodeling by RecG and related eukaryotic enzymes. View Full-Text
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Warren, G.M.; Stein, R.A.; Mchaourab, H.S.; Eichman, B.F. Movement of the RecG Motor Domain upon DNA Binding Is Required for Efficient Fork Reversal. Int. J. Mol. Sci. 2018, 19, 3049.
Warren GM, Stein RA, Mchaourab HS, Eichman BF. Movement of the RecG Motor Domain upon DNA Binding Is Required for Efficient Fork Reversal. International Journal of Molecular Sciences. 2018; 19(10):3049.Chicago/Turabian Style
Warren, Garrett M.; Stein, Richard A.; Mchaourab, Hassane S.; Eichman, Brandt F. 2018. "Movement of the RecG Motor Domain upon DNA Binding Is Required for Efficient Fork Reversal." Int. J. Mol. Sci. 19, no. 10: 3049.
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