{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T15:50:10Z","timestamp":1753890610526,"version":"3.41.2"},"reference-count":28,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T00:00:00Z","timestamp":1662681600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21K19484, 20K08591, 22K06580"],"award-info":[{"award-number":["21K19484, 20K08591, 22K06580"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP20wm0325010"],"award-info":[{"award-number":["JP20wm0325010"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>Irgb6 is a member of interferon \u03b3-induced immunity related GTPase (IRG), and one of twenty \u201ceffector\u201d IRGs, which coordinately attack parasitophorous vacuole membrane (PVM), causing death of intracellular pathogen. Although Irgb6 plays a pivotal role as a pioneer in the process of PVM disruption, the direct effect of Irgb6 on membrane remained to be elucidated. Here, we utilized artificial lipid membranes to reconstitute Irgb6-membrane interaction <jats:italic>in vitro<\/jats:italic>, and revealed that Irgb6 directly deformed the membranes. Liposomes incubated with recombinant Irgb6 were drastically deformed generating massive tubular protrusions in the absence of guanine nucleotide, or with GMP-PNP. Liposome deformation was abolished by incubating with Irgb6-K275A\/R371A, point mutations at membrane targeting residues. The membrane tubules generated by Irgb6 were mostly disappeared by the addition of GTP or GDP, which are caused by detachment of Irgb6 from membrane. Binding of Irgb6 to the membrane, which was reconstituted <jats:italic>in vitro<\/jats:italic> using lipid monolayer, was stimulated at GTP-bound state. Irgb6 GTPase activity was stimulated by the presence of liposomes more than eightfold. Irgb6 GTPase activity in the absence of membrane was also slightly stimulated, by lowering ionic strength, or by increasing protein concentration, indicating synergistic stimulation of the GTPase activity. These results suggest that membrane targeting of Irgb6 and resulting membrane deformation does not require GTP, but converting into GTP-bound state is crucial for detaching Irgb6 from the membrane, which might coincident with local membrane disruption.<\/jats:p>","DOI":"10.3389\/fcimb.2022.992198","type":"journal-article","created":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T19:16:11Z","timestamp":1662750971000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation"],"prefix":"10.3389","volume":"12","author":[{"given":"Hiroshi","family":"Yamada","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tadashi","family":"Abe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hikaru","family":"Nagaoka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eizo","family":"Takashima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryo","family":"Nitta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Yamamoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kohji","family":"Takei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"R92","DOI":"10.1186\/gb-2005-6-11-r92","article-title":"The interferon-inducible p47 (IRG) GTPases in vertebrates: loss of the cell autonomous resistance mechanism in the human lineage","volume":"6","author":"Bekpen","year":"2005","journal-title":"Genome Biol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1038\/nature09032","article-title":"G domain dimerization controls dynamin's assembly-stimulated GTPase activity","volume":"465","author":"Chappie","year":"2010","journal-title":"Nature"},{"key":"B3","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/j.cell.2011.09.003","article-title":"A pseudoatomic model of the dynamin polymer identifies a hydrolysis-dependent powerstroke","volume":"147","author":"Chappie","year":"2011","journal-title":"Cell"},{"key":"B4","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1038\/nature04510","article-title":"How guanylate-binding proteins achieve assembly-stimulated processive cleavage of GTP to GMP","volume":"440","author":"Ghosh","year":"2006","journal-title":"Nature"},{"key":"B5","doi-asserted-by":"publisher","first-page":"727","DOI":"10.1016\/j.molcel.2004.07.017","article-title":"Crystal structure of IIGP1: a paradigm for interferon-inducible p47 resistance GTPases","volume":"15","author":"Ghosh","year":"2004","journal-title":"Mol. Cell"},{"key":"B6","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1038\/s42003-020-01640-7","article-title":"Molecular basis of IRGB10 oligomerization and membrane association for pathogen membrane disruption","volume":"4","author":"Ha","year":"2021","journal-title":"Commun. Biol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"597","DOI":"10.1016\/S0076-6879(05)04052-8","article-title":"In vitro reconstitution of discrete stages of dynamin-dependent endocytosis","volume":"404","author":"Higgins","year":"2005","journal-title":"Methods Enzymol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1016\/j.mib.2011.07.002","article-title":"The IRG protein-based resistance mechanism in mice and its relation to virulence in Toxoplasma gondii","volume":"14","author":"Howard","year":"2011","journal-title":"Curr. Opin. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"2495","DOI":"10.1038\/emboj.2008.176","article-title":"Regulatory interactions between IRG resistance GTPases in the cellular response to Toxoplasma gondii","volume":"27","author":"Hunn","year":"2008","journal-title":"EMBO J."},{"key":"B10","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1111\/j.1462-5822.2010.01443.x","article-title":"Coordinated loading of IRG resistance GTPases on to the Toxoplasma gondii parasitophorous vacuole","volume":"12","author":"Khaminets","year":"2010","journal-title":"Cell Microbiol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"432","DOI":"10.1016\/j.chom.2012.09.007","article-title":"IFN-inducible GTPases in host cell defense","volume":"12","author":"Kim","year":"2012","journal-title":"Cell Host Microbe"},{"key":"B12","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1038\/s41586-018-0378-6","article-title":"Cryo-EM of the dynamin polymer assembled on lipid membrane","volume":"560","author":"Kong","year":"2018","journal-title":"Nature"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.201900549","article-title":"Initial phospholipid-dependent Irgb6 targeting to Toxoplasma gondii vacuoles mediates host defense","volume":"3","author":"Lee","year":"2019","journal-title":"Life Sci. Alliance"},{"key":"B14","doi-asserted-by":"publisher","first-page":"490","DOI":"10.1016\/S0076-6879(05)04043-7","article-title":"Robust colorimetric assays for dynamin's basal and stimulated GTPase activities","volume":"404","author":"Leonard","year":"2005","journal-title":"Methods Enzymol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1038\/nri3210","article-title":"Interferon-inducible effector mechanisms in cell-autonomous immunity","volume":"12","author":"MacMicking","year":"2012","journal-title":"Nat. Rev. Immunol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1111\/cmi.12546","article-title":"Interferon-inducible GTPases in cell autonomous and innate immunity","volume":"18","author":"Meunier","year":"2016","journal-title":"Cell Microbiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/nrm1313","article-title":"The dynamin superfamily: universal membrane tubulation and fission molecules","volume":"5","author":"Praefcke","year":"2004","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1038\/35000617","article-title":"Structure of human guanylate-binding protein 1 representing a unique class of GTP-binding proteins","volume":"403","author":"Prakash","year":"2000","journal-title":"Nature"},{"key":"B19","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.mib.2017.10.021","article-title":"Exposing Toxoplasma gondii hiding inside the vacuole: a role for GBPs, autophagy and host cell death","volume":"40","author":"Saeij","year":"2017","journal-title":"Curr. Opin. Microbiol."},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.202101149","article-title":"Structural basis of membrane recognition of Toxoplasma gondii vacuole by Irgb6","volume":"5","author":"Saijo-Hamano","year":"2021","journal-title":"Life Sci. Alliance"},{"key":"B21","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1034\/j.1600-0854.2000.010503.x","article-title":"Garrotes, springs, ratchets, and whips: putting dynamin models to the test","volume":"1","author":"Sever","year":"2000","journal-title":"Traffic"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.30246","article-title":"Dynamic clustering of dynamin-amphiphysin helices regulates membrane constriction and fission coupled with GTP hydrolysis","volume":"7","author":"Takeda","year":"2018","journal-title":"eLife."},{"key":"B23","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/s0092-8674(00)81228-3","article-title":"Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes","volume":"94","author":"Takei","year":"1998","journal-title":"Cell"},{"key":"B24","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1007\/978-1-60761-447-0_36","article-title":"Use of liposomes to study vesicular transport","volume":"606","author":"Takei","year":"2010","journal-title":"Methods Mol. Biol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"17240","DOI":"10.1016\/S0021-9258(19)85328-0","article-title":"Activation of dynamin GTPase by acidic phospholipids and endogenous rat brain vesicles","volume":"268","author":"Tuma","year":"1993","journal-title":"J. Biol. Chem."},{"key":"B26","doi-asserted-by":"publisher","first-page":"22310","DOI":"10.1074\/jbc.271.37.22310","article-title":"Dynamin self-assembly stimulates its GTPase activity","volume":"271","author":"Warnock","year":"1996","journal-title":"J. Biol. Chem."},{"key":"B27","doi-asserted-by":"publisher","first-page":"302","DOI":"10.1016\/j.immuni.2012.06.009","article-title":"A cluster of interferon-\u03b3-Inducible p65 GTPases plays a critical role in host defense against Toxoplasma gondii","volume":"37","author":"Yamamoto","year":"2012","journal-title":"Immunity"},{"key":"B28","doi-asserted-by":"publisher","first-page":"3483","DOI":"10.1038\/sj.emboj.7600355","article-title":"The stimulatory action of amphiphysin on dynamin function is dependent on lipid bilayer curvature","volume":"23","author":"Yoshida","year":"2004","journal-title":"EMBO J."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2022.992198\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T19:16:16Z","timestamp":1662750976000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2022.992198\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,9]]},"references-count":28,"alternative-id":["10.3389\/fcimb.2022.992198"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2022.992198","relation":{},"ISSN":["2235-2988"],"issn-type":[{"type":"electronic","value":"2235-2988"}],"subject":[],"published":{"date-parts":[[2022,9,9]]},"article-number":"992198"}}