Cellular activities constantly change to precisely respond to their biological needs. In many cases, proteins carry out these activities because they can exhibit graded and dynamic responses to perform an array of cellular functions. To study these biological activities and to repurpose proteins for novel uses such as cell therapies, we must be able to control protein activity with synthetic inducers, such as chemical ligands. Multiple chemical inducers have been employed to achieve protein control, but there remains a need for inducer ligands that minimally interact with endogenous pathways, display high bioavailability, and are absent or minimally present from dietary sources.
In this work, we control protein activities with the Hepatitis C virus cis-protease NS3 and its numerous clinically validated, highly specific inhibitors. First, we use NS3 to create a Ligand Inducible Connection (LInC) to chemically control gene expression, protein localization and cell signaling in mammalian cells. We then extend the use of catalytically inactive NS3 as a high affinity binder in conjunction with genetically encoded approaches to inhibit NS3, including peptides and RNA aptamers. Using catalytically inactive NS3, genetically encoded peptides, and small molecule drugs, we conditionally control peptide docking with antiviral drugs. We apply this concept to control mammalian gene expression, cell signaling, enzyme activity, and develop a new mechanism for allosteric regulation of Cre recombinase. Altogether, we have developed a new toolkit for controlling diverse protein activities with highly orthogonal, antiviral drugs. / 2023-05-15T00:00:00Z
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/42592 |
| Date | 15 May 2021 |
| Creators | Tague, Elliot Parker |
| Contributors | Ngo, John T. |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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