eprintid: 10088353 rev_number: 18 eprint_status: archive userid: 608 dir: disk0/10/08/83/53 datestamp: 2019-12-20 11:46:13 lastmod: 2021-09-17 22:31:42 status_changed: 2019-12-20 11:46:13 type: article metadata_visibility: show creators_name: Benfield, CT creators_name: MacKenzie, F creators_name: Ritzefeld, M creators_name: Mazzon, M creators_name: Weston, S creators_name: Tate, E creators_name: Teo, BH creators_name: Smith, SE creators_name: Kellam, P creators_name: Holmes, EC creators_name: Marsh, M title: Bat IFITM3 restriction depends on S-palmitoylation and a polymorphic site within the CD225 domain ispublished: pub divisions: UCL divisions: B02 divisions: C08 divisions: D77 note: © 2019 Benfield et al. https://creativecommons.org/licenses/by/4.0/ This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0). abstract: Host interferon-induced transmembrane proteins (IFITMs) are broad-spectrum antiviral restriction factors. Of these, IFITM3 potently inhibits viruses that enter cells through acidic endosomes, many of which are zoonotic and emerging viruses with bats (order Chiroptera) as their natural hosts. We previously demonstrated that microbat IFITM3 is antiviral. Here, we show that bat IFITMs are characterized by strong adaptive evolution and identify a highly variable and functionally important site-codon 70-within the conserved CD225 domain of IFITMs. Mutation of this residue in microbat IFITM3 impairs restriction of representatives of four different virus families that enter cells via endosomes. This mutant shows altered subcellular localization and reduced S-palmitoylation, a phenotype copied by mutation of conserved cysteine residues in microbat IFITM3. Furthermore, we show that microbat IFITM3 is S-palmitoylated on cysteine residues C71, C72, and C105, mutation of each cysteine individually impairs virus restriction, and a triple C71A-C72A-C105A mutant loses all restriction activity, concomitant with subcellular re-localization of microbat IFITM3 to Golgi-associated sites. Thus, we propose that S-palmitoylation is critical for Chiropteran IFITM3 function and identify a key molecular determinant of IFITM3 S-palmitoylation. date: 2020-01 date_type: published official_url: https://doi.org/10.26508/lsa.201900542 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1732072 doi: 10.26508/lsa.201900542 pii: 3/1/e201900542 lyricists_name: Marsh, Mark lyricists_name: Mazzon, Michela lyricists_id: MCPMA80 lyricists_id: MMAZZ68 actors_name: Marsh, Mark actors_id: MCPMA80 actors_role: owner full_text_status: public publication: Life Science Alliance volume: 3 number: 1 article_number: e201900542 event_location: United States issn: 2575-1077 citation: Benfield, CT; MacKenzie, F; Ritzefeld, M; Mazzon, M; Weston, S; Tate, E; Teo, BH; ... Marsh, M; + view all <#> Benfield, CT; MacKenzie, F; Ritzefeld, M; Mazzon, M; Weston, S; Tate, E; Teo, BH; Smith, SE; Kellam, P; Holmes, EC; Marsh, M; - view fewer <#> (2020) Bat IFITM3 restriction depends on S-palmitoylation and a polymorphic site within the CD225 domain. Life Science Alliance , 3 (1) , Article e201900542. 10.26508/lsa.201900542 <https://doi.org/10.26508/lsa.201900542>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10088353/1/Benfield%20et%20al.pdf