Molecular basis of ParA ATPase activation by the CTPase ParB during bacterial chromosome segregation

Schnabel, L.; Osorio-Valeriano, M.; Perez-Borrajero, C.; Steinchen, W.; Mais, C.-N.; Simon, B.; Hanssmann, J.; Thamm, M.; Hennig, J.; Bange, G.; Thanbichler, M.

doi:10.1101/2025.03.14.642875

bioRxiv · 10.1101/2025.03.14.642875

Molecular basis of ParA ATPase activation by the CTPase ParB during bacterial chromosome segregation

Schnabel, L.·Osorio-Valeriano, M.·Perez-Borrajero, C.·Steinchen, W.·Mais, C.-N.·Simon, B.·Hanssmann, J.·Thamm, M.·Hennig, J.·Bange, G.·Thanbichler, M.

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Abstract

DNA segregation by bacterial ParABS systems is mediated by transient tethering interactions between nucleoid-bound dimers of the ATPase ParA and centromere (parS)-associated complexes of the clamp-forming CTPase ParB. The lifetime of these interactions is limited by the ParB-dependent activation of ParA ATPase activity. Here, we elucidate the functional interplay between ParA and ParB in the model bacterium Myxococcus xanthus. We demonstrate that the N-terminal ParA-binding motif of ParB associates with a conserved bipartite binding pocket at the ParA dimer interface, in a manner dependent on ParB clamp closure. Moreover, we show that ParB and non-specific DNA interact cooperatively with ParA and synergistically induce structural changes at its Walker A and Walker B motifs that correlate with the activation of ParA ATPase activity. These results advance our understanding of the mechanism underlying DNA transport by the ParABS system and may help to unravel the mode of action of related cargo-positioning systems.

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Schnabel, L., Osorio-Valeriano, M., Perez-Borrajero, C., Steinchen, W., Mais, C.-N., Simon, B., Hanssmann, J., Thamm, M., Hennig, J., Bange, G., Thanbichler, M.. 2025-03-14. Molecular basis of ParA ATPase activation by the CTPase ParB during bacterial chromosome segregation. https://doi.org/10.1101/2025.03.14.642875

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