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bioRxiv · 10.1101/2021.09.23.461529

Nanopore tweezers measurements of RecQ conformational changes reveal the energy landscape of helicase motion

Craig, J. M.·Mills, M.·Laszlo, A.·Kim, H. C.·Abell, S. J.·Huang, J. R.·Mount, J. W.·Neuman, K. C.·Gundlach, J. H.
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Abstract

Helicases are essential for nearly all nucleic acid processes across the tree of life. Using Nanopore Tweezers we observed the small, fast steps taken by single RecQ helicases as they step along and unwind DNA at ultrahigh spatiotemporal resolution. By directly measuring conformational substates of RecQ we determine the coupling between helicase domain motions and chemical reactions that together produce forward motion along the DNA. Application of assisting and opposing forces shows that RecQ has a highly asymmetric energy landscape that reduces its sensitivity to opposing mechanical forces that could be encountered in vivo by molecular roadblocks such as DNA bound proteins. This energy landscape enables RecQ to maintain speed against an opposing load.

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Craig, J. M., Mills, M., Laszlo, A., Kim, H. C., Abell, S. J., Huang, J. R., Mount, J. W., Neuman, K. C., Gundlach, J. H.. 2021-09-23. Nanopore tweezers measurements of RecQ conformational changes reveal the energy landscape of helicase motion. https://doi.org/10.1101/2021.09.23.461529

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