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Consequently, such knowledge of one of the severe acute respiratory syndrome coronavirus 2 (SARS\u2010CoV\u20102)'s proteins is advantageous since the virus is the etiological agent behind one of the biggest health crises of recent times. To that end, we constructed an exhaustive library of bacteria differing from each other by the mutated version of the virus's ORF3a viroporin they harbor. Since the protein is harmful to bacterial growth due to its channel activity, genetic selection followed by deep sequencing could readily identify mutations that abolish the protein's function. Our results have yielded numerous mutations dispersed throughout the sequence that counteract ORF3a's ability to slow bacterial growth. Comparing these data with the conservation pattern of ORF3a within the coronavirinae provided interesting insights: Deleterious mutations obtained in our study corresponded to conserved residues in the protein. However, despite the comprehensive nature of our mutagenesis coverage (108 average mutations per site), we could not reveal all of the protein's conserved residues. Therefore, it is tempting to speculate that our study unearthed positions in the protein pertinent to channel activity, while other conserved residues may correspond to different functionalities of ORF3a. In conclusion, our study provides important information on a key component of SARS\u2010CoV\u20102 and establishes a procedure to analyze other viroporins comprehensively.<\/jats:p>","DOI":"10.1002\/pro.4528","type":"journal-article","created":{"date-parts":[[2022,12,5]],"date-time":"2022-12-05T11:43:28Z","timestamp":1670240608000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Exhaustive mutational analysis of severe acute respiratory syndrome coronavirus 2 <scp>ORF3a<\/scp>: An essential component in the pathogen's infectivity cycle"],"prefix":"10.1002","volume":"32","author":[{"given":"Amit","family":"Benazraf","sequence":"first","affiliation":[{"name":"Department of Biological Chemistry The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem  Jerusalem Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7659-1746","authenticated-orcid":false,"given":"Isaiah T.","family":"Arkin","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem  Jerusalem Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,12,28]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2016.08.007"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2010.08.021"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.369595"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2020.12.142"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btq281"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-020-0771-4"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.113.154963"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/v12080844"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-395-X:3"},{"issue":"1","key":"e_1_2_8_11_1","first-page":"157","article-title":"WHO declares COVID\u201019 a pandemic","volume":"91","author":"Cucinotta D","year":"2020","journal-title":"Acta Biomed"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.144.3620.862"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/148.4.1667"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01695-07"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.10.7108-7115.1996"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1492"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(03)00780-4"},{"key":"e_1_2_8_18_1","unstructured":"GordonA HannonG Fastx\u2010toolkit. 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