title: Tunable and reversible drug control of protein production via a self-excising degron.
abstract: An effective method for direct chemical control over the production of specific proteins would be widely useful. We describe small molecule-assisted shutoff (SMASh), a technique in which proteins are fused to a degron that removes itself in the absence of drug, resulting in the production of an untagged protein. Clinically tested HCV protease inhibitors can then block degron removal, inducing rapid degradation of subsequently synthesized copies of the protein. SMASh allows reversible and dose-dependent shutoff of various proteins in multiple mammalian cell types and in yeast. We also used SMASh to confer drug responsiveness onto an RNA virus for which no licensed inhibitors exist. As SMASh does not require the permanent fusion of a large domain, it should be useful when control over protein production with minimal structural modification is desired. Furthermore, as SMASh involves only a single genetic modification and does not rely on modulating protein-protein interactions, it should be easy to generalize to multiple biological contexts.
authors.affiliation: Department of Biology, Stanford University, Stanford, California, USA.
authors.forename: Hokyung K
authors.initials: HK
authors.lastname: Chung
authors.affiliation: Department of Biology, Stanford University, Stanford, California, USA.
authors.forename: Conor L
authors.initials: CL
authors.lastname: Jacobs
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Yunwen
authors.initials: Y
authors.lastname: Huo
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla, California, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Pediatrics, Emory University, Atlanta, Georgia, USA.
authors.forename: Stefanie A
authors.initials: SA
authors.lastname: Krumm
authors.affiliation: 1] Department of Pediatrics, Emory University, Atlanta, Georgia, USA. [2] Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USA.
authors.forename: Richard K
authors.initials: RK
authors.lastname: Plemper
authors.affiliation: 1] Department of Pharmacology, University of California, San Diego, La Jolla, California, USA. [2] Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA. [3] Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California, USA.
authors.forename: Roger Y
authors.initials: RY
authors.lastname: Tsien
authors.affiliation: 1] Department of Pediatrics, Stanford University, Stanford, California, USA. [2] Department of Bioengineering, Stanford University, Stanford, California, USA.
authors.forename: Michael Z
authors.initials: MZ
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006367:HeLa Cells; D016174:Hepacivirus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007546:Isoquinolines / Q000494:pharmacology*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000821:virology; D061251:Primary Cell Culture; D011480:Protease Inhibitors / Q000494:pharmacology*; D011485:Protein Binding; D059748:Proteolysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology*; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D007546:Isoquinolines; D008164:Luminescent Proteins; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins; D013449:Sulfonamides; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C455041:NS4A cofactor peptide, Hepatitis C virus; C054562:yellow fluorescent protein, Bacteria; C571889:asunaprevir