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Understanding viral pathogenesis at the molecular level is critical to develop effective therapy. Some recent studies have explored SARS\u2010CoV\u20102\u2013host interactomes and provided great resources for understanding viral replication. However, host proteins that functionally associate with SARS\u2010CoV\u20102 are localized in the corresponding subnetwork within the comprehensive human interactome. Therefore, constructing a downstream network including all potential viral receptors, host cell proteases, and cofactors is necessary and should be used as an additional criterion for the validation of critical host machineries used for viral processing. This study applied both affinity purification mass spectrometry (AP\u2010MS) and the complementary proximity\u2010based labeling MS method (BioID\u2010MS) on 29 viral ORFs and 18 host proteins with potential roles in viral replication to map the interactions relevant to viral processing. The analysis yields a list of 693 hub proteins sharing interactions with both viral baits and host baits and revealed their biological significance for SARS\u2010CoV\u20102. Those hub proteins then served as a rational resource for drug repurposing via a virtual screening approach. The overall process resulted in the suggested repurposing of 59 compounds for 15 protein targets. Furthermore, antiviral effects of some candidate drugs were observed\n                    <jats:italic>in\u00a0vitro<\/jats:italic>\n                    validation using image\u2010based drug screen with infectious SARS\u2010CoV\u20102. In addition, our results suggest that the antiviral activity of methotrexate could be associated with its inhibitory effect on specific protein\u2013protein interactions.\n                  <\/jats:p>","DOI":"10.15252\/msb.202110396","type":"journal-article","created":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T10:32:00Z","timestamp":1635417120000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":109,"title":["SARS\u2010CoV\u20102\u2013host proteome interactions for antiviral drug discovery"],"prefix":"10.1038","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9600-0536","authenticated-orcid":false,"given":"Xiaonan","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3857-1805","authenticated-orcid":false,"given":"Sini","family":"Huuskonen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1539-2142","authenticated-orcid":false,"given":"Tuomo","family":"Laitinen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6724-9025","authenticated-orcid":false,"given":"Taras","family":"Redchuk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6608-1120","authenticated-orcid":false,"given":"Mariia","family":"Bogacheva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4471-6698","authenticated-orcid":false,"given":"Kari","family":"Salokas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2801-8902","authenticated-orcid":false,"given":"Ina","family":"P\u00f6hner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6647-8873","authenticated-orcid":false,"given":"Tiina","family":"\u00d6hman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6986-4198","authenticated-orcid":false,"given":"Arun Kumar","family":"Tonduru","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9491-2868","authenticated-orcid":false,"given":"Antti","family":"Hassinen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1950-4749","authenticated-orcid":false,"given":"Lisa","family":"Gawriyski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5555-1975","authenticated-orcid":false,"given":"Salla","family":"Keskitalo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2017-0475","authenticated-orcid":false,"given":"Maria K","family":"Vartiainen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3125-2406","authenticated-orcid":false,"given":"Vilja","family":"Pieti\u00e4inen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4196-4204","authenticated-orcid":false,"given":"Antti","family":"Poso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1340-9732","authenticated-orcid":false,"given":"Markku","family":"Varjosalo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,10,28]]},"reference":[{"key":"BFMSB202110396_CR1","doi-asserted-by":"crossref","first-page":"5066","DOI":"10.3390\/v7092862","volume":"7","author":"M Alenquer","year":"2015","unstructured":"Alenquer M, Amorim MJ (2015) Exosome biogenesis, regulation, and function in viral infection. 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