Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.26.221812v1?rss=1 Authors: Kim, S., Zuromski, K. L., Bell, T. A., Sauer, R. T., Baker, T. A. Abstract: AAA+ proteases, which perform regulated protein degradation in all kingdoms of life, consist of a hexameric AAA+ unfoldase/translocase in complex with a self-compartmentalized peptidase. Based on asymmetric features of cryo-EM structures and a sequential hand-over-hand model of substrate translocation, recent publications have proposed that the AAA+ unfoldases ClpA and ClpX must rotate with respect to their partner peptidase ClpP to allow function. Here, we test this model by covalently crosslinking ClpA to ClpP to prevent rotation. We find that crosslinked ClpAP complexes unfold, translocate, and degrade protein substrates, albeit modestly slower than uncrosslinked enzyme controls. Rotation of ClpA with respect to ClpP therefore is not required for ClpAP protease activity, although some flexibility in how the AAA+ ring docks on ClpP may be necessary for optimal function. Copy rights belong to original authors. Visit the link for more info