{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T15:59:37Z","timestamp":1790438377575,"version":"4.1.0"},"reference-count":39,"publisher":"National Academy of Sciences","issue":"19","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2006,5,9]]},"abstract":"<jats:p>\n                    VirB8-like proteins are essential components of type IV secretion systems, bacterial virulence factors that mediate the translocation of effector molecules from many bacterial pathogens into eukaryotic cells. Based on cell biological, genetic, and x-ray crystallographic data, VirB8 was proposed to undergo multiple protein\u2013protein interactions to mediate assembly of the translocation machinery. Here we report the results of a structure\u2013function analysis of the periplasmic domain of VirB8 from the mammalian pathogen\n                    <jats:italic>Brucella suis<\/jats:italic>\n                    , which identifies amino acid residues required for three protein\u2013protein interactions. VirB8 variants changed at residues proposed to be involved in dimerization, and protein\u2013protein interactions were purified and characterized\n                    <jats:italic>in vitro<\/jats:italic>\n                    and\n                    <jats:italic>in vivo<\/jats:italic>\n                    . Changes at M102, Y105, and E214 affected the self-association as measured by analytical ultracentrifugation and gel filtration. The interaction with\n                    <jats:italic>B. suis<\/jats:italic>\n                    VirB10 was reduced by changes at T201, and change at R230 inhibited the interaction with VirB4\n                    <jats:italic>in vitro<\/jats:italic>\n                    . The\n                    <jats:italic>in vivo<\/jats:italic>\n                    functionality of VirB8 variants was determined by complementation of growth in macrophages by a\n                    <jats:italic>B. suis virB8<\/jats:italic>\n                    mutant and by using a heterologous assay of type IV secretion system assembly in\n                    <jats:italic>Agrobacterium tumefaciens<\/jats:italic>\n                    . Changes at Y105, T201, R230, and at several other residues impaired the\n                    <jats:italic>in vivo<\/jats:italic>\n                    function of VirB8, suggesting that we have identified interaction sites of relevance in the natural biological context.\n                  <\/jats:p>","DOI":"10.1073\/pnas.0600862103","type":"journal-article","created":{"date-parts":[[2006,4,28]],"date-time":"2006-04-28T20:39:51Z","timestamp":1146256791000},"page":"7252-7257","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":52,"title":["Dimerization and interactions of\n                    <i>Brucella suis<\/i>\n                    VirB8 with VirB4 and VirB10 are required for its biological activity"],"prefix":"10.1073","volume":"103","author":[{"given":"Athanasios","family":"Paschos","sequence":"first","affiliation":[{"name":"*Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 4K1;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilles","family":"Patey","sequence":"additional","affiliation":[{"name":"Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale U431, Facult\u00e9 de M\u00e9decine, Avenue Kennedy, F-30900 N\u00eemes, France;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Durga","family":"Sivanesan","sequence":"additional","affiliation":[{"name":"*Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 4K1;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chan","family":"Gao","sequence":"additional","affiliation":[{"name":"*Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 4K1;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard","family":"Bayliss","sequence":"additional","affiliation":[{"name":"School of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom;"},{"name":"Institute of Structural Molecular Biology, University College London\/Birkbeck, Malet Street, London WC1E 7HX, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Waksman","sequence":"additional","affiliation":[{"name":"School of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom;"},{"name":"Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom; and"},{"name":"Institute of Structural Molecular Biology, University College London\/Birkbeck, Malet Street, London WC1E 7HX, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"O'Callaghan","sequence":"additional","affiliation":[{"name":"Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale U431, Facult\u00e9 de M\u00e9decine, Avenue Kennedy, F-30900 N\u00eemes, France;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Baron","sequence":"additional","affiliation":[{"name":"*Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 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