Chromatinization Modulates Topoisomerase II Processivity

Lee, J.; Wu, M.; Inman, J.; Singh, G.; Park, S. h.; Lee, J. H.; Fulbright, R. M.; Hong, Y.; Jeong, J. D.; Berger, J. M.; Wang, M. D.

doi:10.1101/2023.10.03.560726

bioRxiv · 10.1101/2023.10.03.560726

Chromatinization Modulates Topoisomerase II Processivity

Lee, J.·Wu, M.·Inman, J.·Singh, G.·Park, S. h.·Lee, J. H.·Fulbright, R. M.·Hong, Y.·Jeong, J. D.·Berger, J. M.·Wang, M. D.

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Abstract

Type IIA topoisomerases are essential DNA processing enzymes that must robustly and reliably relax DNA torsional stress in vivo. While cellular processes constantly create different degrees of torsional stress, how this stress feeds back to control type IIA topoisomerase function remains obscure. Using a suite of single-molecule approaches, we examined the torsional impact on supercoiling relaxation of both naked DNA and chromatin by eukaryotic topoisomerase II (topo II). We observed that topo II was at least [~] 50-fold more processive on plectonemic DNA than previously estimated, capable of relaxing > 6000 turns. We further discovered that topo II could relax supercoiled DNA prior to plectoneme formation, but with a [~]100-fold reduction in processivity; strikingly, the relaxation rate in this regime decreased with diminishing torsion in a manner consistent with the capture of transient DNA loops by topo II. Chromatinization preserved the high processivity of the enzyme under high torsional stress. Interestingly, topo II was still highly processive ([~] 1000 turns) even under low torsional stress, consistent with the predisposition of chromatin to readily form DNA crossings. This work establishes that chromatin is a major stimulant of topo II function, capable of enhancing function even under low torsional stress.

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Lee, J., Wu, M., Inman, J., Singh, G., Park, S. h., Lee, J. H., Fulbright, R. M., Hong, Y., Jeong, J. D., Berger, J. M., Wang, M. D.. 2023-10-04. Chromatinization Modulates Topoisomerase II Processivity. https://doi.org/10.1101/2023.10.03.560726

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