DNA gyrase in live bacteria forms liquid condensates through weak multivalent bonding of excess GyrB

Syeda, A.; Hollands, K.; Frame, L.; Shepherd, J.; Payne-Dwyer, A.; Goffee, E.; Burton, N.; Basu, A.; Noy, A.; Maxwell, A.; Leake, M. C.

doi:10.64898/2026.08.13.744598

bioRxiv · 10.64898/2026.08.13.744598

DNA gyrase in live bacteria forms liquid condensates through weak multivalent bonding of excess GyrB

Syeda, A.·Hollands, K.·Frame, L.·Shepherd, J.·Payne-Dwyer, A.·Goffee, E.·Burton, N.·Basu, A.·Noy, A.·Maxwell, A.·Leake, M. C.

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Abstract

Type IIA bacterial topoisomerase DNA gyrase, a GyrA/GyrB heterotetramer, has crucial roles maintaining transcription and DNA replication by relaxing positive DNA supercoils through introducing negative supercoils. However, rates of gyrase-catalysed supercoiling in vitro cannot explain much higher rates required in vivo. To address this puzzle, we used high-speed single-molecule fluorescence imaging of GyrA/GyrB reporters in live Escherichia coli, indicating that cells contain [~]40% more GyrB than GyrA expressed in a diffuse pool or in clusters whose mobility depends on whether they are bound to DNA. Unexpectedly, we discovered that clusters are non-stoichiometric containing [~]150% more GyrB than GyrA, significantly greater than the cellular average, with fluorescence recovery after photobleaching revealing that clustered GyrA and GyrB behave as a liquid whose abundance can be increased by applying gyrase-targeting antibiotics. Structural docking indicates that the liquid state is stabilised through excess GyrB progressively binding to existing clusters via weak, multivalent interactions. By operating in liquid condensates, A2B2 that dissociates from DNA can rebind rapidly instead of diffusing away, increasing enzyme processivity to enable multiple rounds of catalysis that can keep pace with transcription and DNA replication in vivo. This demonstrates a new role for condensates of overcoming kinetic limitations imposed by diffusion. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=158 SRC="FIGDIR/small/744598v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@1514d4aorg.highwire.dtl.DTLVardef@a95ceeorg.highwire.dtl.DTLVardef@11417daorg.highwire.dtl.DTLVardef@2a2856_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Syeda, A., Hollands, K., Frame, L., Shepherd, J., Payne-Dwyer, A., Goffee, E., Burton, N., Basu, A., Noy, A., Maxwell, A., Leake, M. C.. 2026-08-17. DNA gyrase in live bacteria forms liquid condensates through weak multivalent bonding of excess GyrB. https://doi.org/10.64898/2026.08.13.744598

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