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.
","abstract_html":"<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>","abstract_has_math":false,"creators":["Kilianski, Andrew"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation","degree_discipline":"Microbiology and Immunology","degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-01-01T08:00:00Z","date_published":"2014-01-01T08:00:00Z","updated_at":"2026-07-24T02:56:24Z","subjects":["coronavirus","MERS","NFkB","PLP","protease","SARS","Virology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://ecommons.luc.edu/luc_diss/1272","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Kilianski, Andrew"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2016-03-31T17:20:35Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology and Immunology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["coronavirus","MERS","NFkB","PLP","protease","SARS","Virology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://ecommons.luc.edu/luc_diss/1272"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.
"]},{"key":"dc:title","label":"Title","values":["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"]}]}],"canonical_facts":{"dc:creator":["Kilianski, Andrew"],"dc:date.available":["2016-03-31T17:20:35Z"],"dc:description.abstract":["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.
"],"dc:identifier":["https://ecommons.luc.edu/luc_diss/1272"],"dc:subject":["coronavirus","MERS","NFkB","PLP","protease","SARS","Virology"],"dc:title":["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"],"thesis:degree_discipline":["Microbiology and Immunology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T02:56:24Z"}