Controlled protein activities with viral proteases, antiviral peptides, and antiviral drugs

Tague, E. P.; McMahan, J. B.; Tague, N.; Dunlop, M. J.; Ngo, J. T.

doi:10.1101/2023.02.27.530290

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bioRxiv · 10.1101/2023.02.27.530290

Controlled protein activities with viral proteases, antiviral peptides, and antiviral drugs

Tague, E. P.·McMahan, J. B.·Tague, N.·Dunlop, M. J.·Ngo, J. T.

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Abstract

Chemical control of protein activity is a powerful tool for scientific study, synthetic biology, and cell therapy; however, for broad use, effective chemical inducer systems must minimally crosstalk with endogenous processes and exhibit desirable drug delivery properties. Accordingly, the drug-controllable proteolytic activity of hepatitis C cis-protease NS3 and its associated antiviral drugs have been used to regulate protein activity and gene modulation. These tools advantageously exploit non-eukaryotic/prokaryotic proteins and clinically approved inhibitors. Here we expand the toolkit by utilizing catalytically inactive NS3 protease as a high affinity binder to genetically encoded, antiviral peptides. Through our approach, we create NS3-peptide complexes that can be displaced by FDA-approved drugs to modulate transcription, cell signaling, split-protein complementation. With our developed system, we discover a new mechanism to allosterically regulate Cre recombinase. Allosteric Cre regulation with NS3 ligands enables orthogonal recombination tools in eukaryotic cells and functions in divergent organisms to control prokaryotic recombinase activity.

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Tague, E. P., McMahan, J. B., Tague, N., Dunlop, M. J., Ngo, J. T.. 2023-02-27. Controlled protein activities with viral proteases, antiviral peptides, and antiviral drugs. https://doi.org/10.1101/2023.02.27.530290

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