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Here, we aimed at better characterizing the cellular responses induced by these proteins. First, we show that expression of SARS-CoV-2 E or 3a protein in CHO cells gives rise to cells with newly acquired round shapes that detach from the Petri dish. This suggests that cell death is induced upon expression of E or 3a protein. We confirmed this by using flow cytometry. In adhering cells expressing E or 3a protein, the whole-cell currents were not different from those of the control, suggesting that E and 3a proteins are not plasma membrane viroporins. In contrast, recording the currents on detached cells uncovered outwardly rectifying currents much larger than those observed in the control. We illustrate for the first time that carbenoxolone and probenecid block these outwardly rectifying currents; thus, these currents are most probably conducted by pannexin channels that are activated by cell morphology changes and also potentially by cell death. The truncation of C-terminal PDZ binding motifs reduces the proportion of dying cells but does not prevent these outwardly rectifying currents. This suggests distinct pathways for the induction of these cellular events by the two proteins. We conclude that SARS-CoV-2 E and 3a proteins are not viroporins expressed at the plasma membrane.<\/jats:p>","DOI":"10.3390\/cells12111474","type":"journal-article","created":{"date-parts":[[2023,5,25]],"date-time":"2023-05-25T04:35:54Z","timestamp":1684989354000},"page":"1474","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["SARS-CoV-2 E and 3a Proteins Are Inducers of Pannexin Currents"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2269-5750","authenticated-orcid":false,"given":"Barbara B. R.","family":"Oliveira-Mendes","sequence":"first","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Malak","family":"Alameh","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"},{"name":"Labex Ion Channels, Science and Therapeutics, F-06560 Valbonne, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B\u00e9atrice","family":"Ollivier","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5100-5571","authenticated-orcid":false,"given":"J\u00e9r\u00f4me","family":"Montnach","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicolas","family":"Bid\u00e8re","sequence":"additional","affiliation":[{"name":"Team SOAP, CRCI2NA, INSERM, CNRS, Nantes Universit\u00e9, Universit\u00e9 d\u2019Angers, F-44000 Nantes, France"},{"name":"Equipe Labellis\u00e9e Ligue Contre le Cancer, F-75006 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0476-2603","authenticated-orcid":false,"given":"Fr\u00e9d\u00e9rique","family":"Souaz\u00e9","sequence":"additional","affiliation":[{"name":"CRCI2NA INSERM, CNRS, Nantes Universit\u00e9, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicolas","family":"Escriou","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, D\u00e9partement de Sant\u00e9 Globale, F-75015 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Flavien","family":"Charpentier","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4850-4171","authenticated-orcid":false,"given":"Isabelle","family":"Bar\u00f3","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2782-9615","authenticated-orcid":false,"given":"Michel","family":"De Waard","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"},{"name":"Labex Ion Channels, Science and Therapeutics, F-06560 Valbonne, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8007-5931","authenticated-orcid":false,"given":"Gildas","family":"Loussouarn","sequence":"additional","affiliation":[{"name":"L\u2019institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1967","DOI":"10.1056\/NEJMoa030747","article-title":"Identification of a Novel Coronavirus in Patients with Severe Acute Respiratory Syndrome","volume":"348","author":"Drosten","year":"2003","journal-title":"N. 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