Update: our preprint is now published at eLife (HERE)
A follow-up from great works by Gruber and Bange & Thanbichler labs: we show that Caulobacter ParB (and 8 other chromosomal ParB proteins) requires cytidine triphosphate (CTP) to spread in vitro: https://bit.ly/2rIO0Lh
We show that accumulation of Caulobacter ParB requires a closed DNA substrate with blocked ends and that a DNA-binding transcriptional regulator can act as a roadblock to attenuate spreading unidirectionally in vitro.
Our real-time and label-free in vitro reconstitution of Caulobacter ParB spreading has recapitulated many well-known aspects of ParB behaviors.
The ease and medium-throughput manner of our methodology hopefully will facilitate future works by the community to investigate many other aspects of ParB and bacterial chromosome segregation.
We thank the Gruber and Thanbichler groups for generously sharing bespoke reagents and pre-publication data.