Articles | Volume 2, issue 2
https://doi.org/10.5194/mr-2-715-2021
https://doi.org/10.5194/mr-2-715-2021
Research article
 | 
14 Sep 2021
Research article |  | 14 Sep 2021

Rapid assessment of Watson–Crick to Hoogsteen exchange in unlabeled DNA duplexes using high-power SELOPE imino 1H CEST

Bei Liu, Atul Rangadurai, Honglue Shi, and Hashim M. Al-Hashimi

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Field: Liquid-state NMR | Topic: Applications – biological macromolecules
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Cited articles

Afek, A., Shi, H., Rangadurai, A., Sahay, H., Senitzki, A., Xhani, S., Fang, M., Salinas, R., Mielko, Z., Pufall, M. A., Poon, G. M. K., Haran, T. E., Schumacher, M. A., Al-Hashimi, H. M., and Gordan, R.: DNA mismatches reveal conformational penalties in protein-DNA recognition, Nature, 587, 291–296, https://doi.org/10.1038/s41586-020-2843-2, 2020. 
Aishima, J., Gitti, R. K., Noah, J. E., Gan, H. H., Schlick, T., and Wolberger, C.: A Hoogsteen base pair embedded in undistorted B-DNA, Nucleic Acids Res., 30, 5244–5252, 2002. 
Alvey, H. S., Gottardo, F. L., Nikolova, E. N., and Al-Hashimi, H. M.: Widespread transient Hoogsteen base pairs in canonical duplex DNA with variable energetics, Nat. Commun., 5, 4786, https://doi.org/10.1038/ncomms5786, 2014. 
Baronti, L., Guzzetti, I., Ebrahimi, P., Friebe Sandoz, S., Steiner, E., Schlagnitweit, J., Fromm, B., Silva, L., Fontana, C., Chen, A. A., and Petzold, K.: Base-pair conformational switch modulates miR-34a targeting of Sirt1 mRNA, Nature, 583, 139–144, https://doi.org/10.1038/s41586-020-2336-3, 2020. 
Ben Imeddourene, A., Zargarian, L., Buckle, M., Hartmann, B., and Mauffret, O.: Slow motions in A.T rich DNA sequence, Sci. Rep., 10, 19005, https://doi.org/10.1038/s41598-020-75645-x, 2020. 
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Short summary
There is growing interest in mapping exchange dynamics between Watson–Crick and Hoogsteen conformations across different DNA contexts. However, current methods are ill-suited for measurements at a large scale because they require isotopically enriched samples. We report that Hoogsteen dynamics can be measured on unlabeled samples using 1H CEST experiments, which have higher throughput and lower cost relative to conventional methods and also provide new insights into Hoogsteen dynamics.