Department of Biochemistry and Molecular Pharmacology
Amino Acids, Peptides, and Proteins | Biophysics | Enzymes and Coenzymes
Many viruses utilize ringed packaging ATPases to translocate double-stranded DNA into procapsids during replication. A critical step in the mechanochemical cycle of such ATPases is ATP binding, which causes a subunit within the motor to grip DNA tightly. Here, we probe the underlying molecular mechanism by which ATP binding is coupled to DNA gripping and show that a glutamate switch residue found in AAA+ enzymes is central to this coupling in viral packaging ATPases. Using free energy landscapes computed through molecular dynamics simulations, we determined the stable conformational state of the ATPase active site in apo, ATP-bound, and ADP-bound states. Our results show that the catalytic glutamate residue transitions from an inactive to an active pose upon ATP binding, and that a residue assigned as the glutamate switch is necessary for regulating the transition. Further, we identified via mutual information analyses the intramolecular signaling pathway mediated by the glutamate switch that is responsible for coupling ATP binding to conformational transitions of DNA-gripping motifs. We corroborated these predictions with both structural and functional experimental data. Specifically, we showed that the crystal structure of the ADP-bound P74-26 packaging ATPase is consistent with the predicted structural coupling from simulations, and we further showed that disrupting the predicted signaling pathway indeed decouples ATPase activity from DNA translocation activity in the φ29 DNA packaging motor. Our work thus establishes a signaling pathway in viral DNA packaging motors that ensures coordination between chemical and mechanical events involved in viral DNA packaging.
Biophysics, ATPases, procapsids, viral DNA packaging
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DOI of Published Version
bioRxiv 2020.12.01.406595; doi: https://doi.org/10.1101/2020.12.01.406595. Link to preprint on bioRxiv.
Pajak J, Atz R, Hilbert BJ, Morais MC, Kelch BA, Jardine P. (2020). Viral Packaging ATPases Utilize a Glutamate Switch to Couple ATPase Activity and DNA Translocation [preprint]. UMass Chan Medical School Faculty Publications. https://doi.org/10.1101/2020.12.01.406595. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/1874
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