Coronavirus Proteases as Therapeutic Targets: Development of Biosensors to Detect Inhibition of Protease Activity and Separation of the Multiple Functions of Coronavirus Papain-Like Proteases

<p>Coronaviruses are important human pathogens and have the potential to severely impact public health on an international scale. The emergence of SARS-CoV and MERS-CoV highlight the need for research to identify antivirals and vaccines against coronaviruses. To develop therapeutics against current and potentially emergent coronaviruses, I utilized two approaches targeting the proteases encoded within all coronaviruses. The papain-like protease and 3C-like protease of coronaviruses are responsible for cleaving viral polyproteins early during infection, and this step is required for viral replication. To quantitatively assess the inhibition by small-molecule compounds on MERS-CoV protease activity, I developed a luciferase-based biosensor to monitor protease cleavage within cells. Using this assay, I demonstrated that an inhibitor that is efficacious against SARS-CoV had activity against the 3C-like protease of MERS-CoV. In the second approach, I investigated the multifunctional papain-like protease of SARS-CoV, which has been implicated in pathogenesis by acting as a deubiquitinating (DUB) enzyme and blocking host immune responses. To determine if PLpro DUB activity is responsible for innate immune antagonism, I mutated residues predicted to interact with ubiquitin and discovered that when this interaction was interrupted, PLpro was unable to antagonize innate immune pathways. Engineering these mutations into SARS-COV may generate an attenuated virus that could stimulate a protective immune response in the absence of disease.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology and Immunology

Year dc:date.available

2014

Author dc:creator

• Kilianski, Andrew

• coronavirus
• MERS
• NFkB
• PLP
• protease
• SARS
• Virology

Repository record dc:identifier

https://ecommons.luc.edu/luc_diss/1272

OAI identifier oai:identifier

oai:ecommons.luc.edu:luc_diss-2271