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caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a significant public health burden with limited treatment options. Many \u03b2-coronaviruses, including SARS-CoV-2, gain entry to host cells through the interaction of SARS-CoV-2 spike protein with membrane-bound ACE2 (angiotensin-converting enzyme 2). Given its necessity for SARS-CoV-2 infection, ACE2 represents a potential therapeutic target in COVID-19. However, early attempts focusing on ACE2 in COVID-19 have not validated it as a druggable target nor identified other ACE2-related novel proteins for therapeutic intervention. Here, we identify a mechanism for ACE2 protein modulation by the deubiquitinase (DUB) enzyme UCHL1 (ubiquitin carboxyl-terminal hydrolase isozyme L1). ACE2 is constitutively ubiquitinated and degraded by the proteasome in lung epithelia. SARS-CoV-2 spike protein cellular internalization increased ACE2 protein abundance by decreasing its degradation. Using an siRNA library targeting 96 human DUBs, we identified UCHL1 as a putative regulator of ACE2 function as a viral receptor. Overexpressed UCHL1 preserved ACE2 protein abundance, whereas silencing of the DUB in cells destabilized ACE2 through increased polyubiquitination. A commercially available small molecule inhibitor of UCHL1 DUB activity decreased ACE2 protein concentrations coupled with inhibition of SARS-CoV-2 infection in epithelial cells. These findings describe a unique pathway of ACE2 regulation uncovering UCHL1 as a potential therapeutic target to modulate COVID-19 viral entry as a platform for future small molecule design and testing.<\/jats:p>","DOI":"10.1165\/rcmb.2022-0331oc","type":"journal-article","created":{"date-parts":[[2023,2,2]],"date-time":"2023-02-02T14:52:54Z","timestamp":1675349574000},"page":"566-576","update-policy":"https:\/\/doi.org\/10.1164\/crossmarkpolicies","source":"Crossref","is-referenced-by-count":10,"title":["Inhibiting the Deubiquitinase UCHL1 Reduces SARS-CoV-2 Viral Uptake by ACE2"],"prefix":"10.1093","volume":"68","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3576-4388","authenticated-orcid":false,"given":"Joseph S.","family":"Bednash","sequence":"first","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Finny","family":"Johns","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniela","family":"Farkas","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ajit","family":"Elhance","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica","family":"Adair","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kirstin","family":"Cress","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6128-4575","authenticated-orcid":false,"given":"Jacob S.","family":"Yount","sequence":"additional","affiliation":[{"name":"Department of Microbial Infection and Immunity, The Ohio State University, Columbus, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adam D.","family":"Kenney","sequence":"additional","affiliation":[{"name":"Department of Microbial Infection and Immunity, The Ohio State University, Columbus, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James D.","family":"Londino","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal 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