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Although EVs have been shown to usurp the cellular recycling process of autophagy for pro-viral functions, the precise manner by which this is accomplished remains to be elucidated. In the current manuscript, we sought to address the mechanism by which EVs subvert the autophagy pathway using Coxsackievirus B3 (CVB3) as a model. We showed that CVB3 infection selectively degrades the autophagy cysteine protease ATG4A but not other isoforms. Exogenous expression of an N-terminally Flag-labeled ATG4A demonstrated the emergence of a 43-kDa cleavage fragment following CVB3 infection. Furthermore, bioinformatics analysis coupled with site-directed mutagenesis and in vitro cleavage assays revealed that CVB3 protease 2A cleaves ATG4A before glycine 374. Using a combination of genetic silencing and overexpression studies, we demonstrated a novel pro-viral function for the autophagy protease ATG4A. Additionally, cleavage of ATG4A was associated with a loss of autophagy function of the truncated cleavage fragment. Collectively, our study identified ATG4A as a novel substrate of CVB3 protease, leading to disrupted host cellular function and sheds further light on viral mechanisms of autophagy dysregulation.<\/jats:p>","DOI":"10.3390\/v14092026","type":"journal-article","created":{"date-parts":[[2022,9,14]],"date-time":"2022-09-14T00:15:02Z","timestamp":1663114502000},"page":"2026","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Coxsackievirus Protease 2A Targets Host Protease ATG4A to Impair Autophagy"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4614-665X","authenticated-orcid":false,"given":"Yiyun Michelle","family":"Fan","sequence":"first","affiliation":[{"name":"Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC V6T 1Z3, Canada"},{"name":"Centre for Heart Lung Innovation, St. Paul\u2019s Hospital, Vancouver, BC V6Z 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yizhuo Lyanne","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC V6T 1Z3, Canada"},{"name":"Centre for Heart Lung Innovation, St. Paul\u2019s Hospital, Vancouver, BC V6Z 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amirhossein","family":"Bahreyni","sequence":"additional","affiliation":[{"name":"Centre for Heart Lung Innovation, St. Paul\u2019s Hospital, Vancouver, BC V6Z 1Y6, Canada"},{"name":"Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 2B5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7708-7840","authenticated-orcid":false,"given":"Honglin","family":"Luo","sequence":"additional","affiliation":[{"name":"Centre for Heart Lung Innovation, St. Paul\u2019s Hospital, Vancouver, BC V6Z 1Y6, Canada"},{"name":"Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 2B5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3950-2070","authenticated-orcid":false,"given":"Yasir","family":"Mohamud","sequence":"additional","affiliation":[{"name":"Centre for Heart Lung Innovation, St. Paul\u2019s Hospital, Vancouver, BC V6Z 1Y6, Canada"},{"name":"Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 2B5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1080\/21505594.2018.1551010","article-title":"The Intertwined Life Cycles of Enterovirus and Autophagy","volume":"10","author":"Mohamud","year":"2019","journal-title":"Virulence"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"629","DOI":"10.2217\/fmb.15.5","article-title":"Coxsackievirus B3 replication and pathogenesis","volume":"10","author":"Garmaroudi","year":"2015","journal-title":"Future Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1161\/CIRCRESAHA.115.306573","article-title":"Myocarditis","volume":"118","author":"Fung","year":"2016","journal-title":"Circ. Res."},{"key":"ref_4","first-page":"1","article-title":"Falling Short: How Canada is Failing People with Heart Failure","volume":"1","author":"Canada","year":"2022","journal-title":"Spotlight Heart Fail."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1038\/s41579-018-0022-3","article-title":"The life cycle of non-polio enteroviruses and how to target it (vol 16, pg 368, 2018)","volume":"16","author":"Baggen","year":"2018","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2861","DOI":"10.1101\/gad.1599207","article-title":"Autophagy: Process and function","volume":"21","author":"Mizushima","year":"2007","journal-title":"Genes Dev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/path.2697","article-title":"Autophagy: Cellular and molecular mechanisms","volume":"221","author":"Glick","year":"2010","journal-title":"J. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1089\/ars.2013.5371","article-title":"An overview of autophagy: Morphology, mechanism, and regulation","volume":"20","author":"Parzych","year":"2014","journal-title":"Antioxid. Redox Signal."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2020.1797280","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)","volume":"17","author":"Klionsky","year":"2021","journal-title":"Autophagy"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"424","DOI":"10.5483\/BMBRep.2016.49.8.081","article-title":"Role of the mammalian ATG8\/LC3 family in autophagy: Differential and compensatory roles in the spatiotemporal regulation of autophagy","volume":"49","author":"Lee","year":"2016","journal-title":"BMB Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"883","DOI":"10.4161\/auto.19652","article-title":"Dual roles of Atg8-PE deconjugation by Atg4 in autophagy","volume":"8","author":"Yu","year":"2012","journal-title":"Autophagy"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1038\/ja.2017.104","article-title":"Autophagy-regulating protease Atg4: Structure, function, regulation and inhibition","volume":"71","author":"Maruyama","year":"2018","journal-title":"J. Antibiot."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"7327","DOI":"10.1074\/jbc.M110.199059","article-title":"Kinetics comparisons of mammalian Atg4 homologues indicate selective preferences toward diverse Atg8 substrates","volume":"286","author":"Li","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.virol.2015.03.042","article-title":"Viruses and the autophagy pathway","volume":"479","author":"Jackson","year":"2015","journal-title":"Virology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.semcdb.2019.08.001","article-title":"The interplay of autophagy and enterovirus","volume":"101","author":"Huang","year":"2020","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1080\/15548627.2018.1458805","article-title":"Poliovirus induces autophagic signaling independent of the ULK1 complex","volume":"14","author":"Velazquez","year":"2018","journal-title":"Autophagy"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"19068","DOI":"10.1038\/s41598-020-76227-7","article-title":"Coxsackievirus infection induces a non-canonical autophagy independent of the ULK and PI3K complexes","volume":"10","author":"Mohamud","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Xiang, P., Mohamud, Y., and Luo, H. (2021). SNAP47 Interacts with ATG14 to Promote VP1 Conjugation and CVB3 Propagation. Cells, 10.","DOI":"10.3390\/cells10082141"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3924","DOI":"10.1080\/15548627.2021.1896925","article-title":"Coxsackievirus B3 targets TFEB to disrupt lysosomal function","volume":"17","author":"Mohamud","year":"2021","journal-title":"Autophagy"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/bs.mie.2016.10.024","article-title":"Measurement of the Activity of the Atg4 Cysteine Proteases","volume":"587","author":"Li","year":"2017","journal-title":"Method Enzymol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3292","DOI":"10.1016\/j.celrep.2018.02.090","article-title":"Enteroviral Infection Inhibits Autophagic Flux via Disruption of the SNARE Complex to Enhance Viral Replication","volume":"22","author":"Mohamud","year":"2018","journal-title":"Cell Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"8444","DOI":"10.1128\/jvi.70.12.8444-8450.1996","article-title":"The eIF4G-eIF4E complex is the target for direct cleavage by the rhinovirus 2A proteinase","volume":"70","author":"Haghighat","year":"1996","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1016\/j.virol.2010.12.014","article-title":"Enterovirus infections of the central nervous system","volume":"411","author":"Rhoades","year":"2011","journal-title":"Virology"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Saeed, M., Kapell, S., Hertz, N.T., Wu, X.F., Bell, K., Ashbrook, A.W., Mark, M.T., Zebroski, H.A., Neal, M.L., and Flodstrom-Tullberg, M. (2020). Defining the proteolytic landscape during enterovirus infection. PLoS Pathog., 16.","DOI":"10.1371\/journal.ppat.1008927"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e02211-17","DOI":"10.1128\/JVI.02211-17","article-title":"N-Terminomics TAILS Identifies Host Cell Substrates of Poliovirus and Coxsackievirus B3 3C Proteinases That Modulate Virus Infection","volume":"92","author":"Jagdeo","year":"2018","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"eabc2691","DOI":"10.1126\/sciimmunol.abc2691","article-title":"Defects in LC3B2 and ATG4A underlie HSV2 meningitis and reveal a critical role for autophagy in antiviral defense in humans","volume":"5","author":"Hait","year":"2020","journal-title":"Sci. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1817","DOI":"10.3389\/fmicb.2020.01817","article-title":"Enterovirus Replication Organelles and Inhibitors of Their Formation","volume":"11","author":"Li","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9143","DOI":"10.1128\/JVI.00641-08","article-title":"Autophagosome supports coxsackievirus B3 replication in host cells","volume":"82","author":"Wong","year":"2008","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1128\/JVI.05937-11","article-title":"Complex Dynamic Development of Poliovirus Membranous Replication Complexes","volume":"86","author":"Belov","year":"2012","journal-title":"J. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3304","DOI":"10.1016\/j.celrep.2018.03.003","article-title":"Enteroviruses Remodel Autophagic Trafficking through Regulation of Host SNARE Proteins to Promote Virus Replication and Cell Exit","volume":"22","author":"Corona","year":"2018","journal-title":"Cell Rep."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/9\/2026\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:30:40Z","timestamp":1760142640000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/9\/2026"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,13]]},"references-count":30,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["v14092026"],"URL":"https:\/\/doi.org\/10.3390\/v14092026","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,13]]}}}