{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T18:51:15Z","timestamp":1789930275479,"version":"4.0.1"},"reference-count":49,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2009,5,1]],"date-time":"2009-05-01T00:00:00Z","timestamp":1241136000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2009,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>The proteinVirB8 plays a critical role in the assembly and function of the<jats:italic>Agrobacterium tumefaciens virB<\/jats:italic>type IV secretion system (T4SS). The structure of the periplasmic domain of both<jats:italic>A<\/jats:italic>.<jats:italic>tumefaciens<\/jats:italic>and<jats:italic>Brucella suis<\/jats:italic>VirB8 has been determined, and site-directed mutagenesis has revealed amino acids involved in the dimerization of VirB8 and interactions with VirB4 and VirB10. We have shown previously that TraJ, the VirB8 homologue from pSB102, and the chimeric protein TraJB8, encompassing the cytoplasmic and transmembrane (TM) domains of TraJ and the periplasmic domain of VirB8, were unable to complement a<jats:italic>B<\/jats:italic>.<jats:italic>suis<\/jats:italic>mutant containing an in-frame deletion of the<jats:italic>virB8<\/jats:italic>gene. This suggested that the presence of the TraJ cytoplasmic and TM domains could block VirB8 dimerization or assembly in the inner membrane. By bacterial two-hybrid analysis, we found that VirB8, TraJ, and the chimeras can all interact to form both homo- and heterodimers. However, the presence of the TM domain of TraJ resulted in much stronger interactions in both the homo- and heterodimers. We expressed the wild-type and chimeric proteins in wild-type<jats:italic>B<\/jats:italic>.<jats:italic>suis<\/jats:italic>. The presence of proteins carrying the TM domain of TraJ had a dominant negative effect, leading to complete loss of virulence. This suggests that the T4SS is a dynamic structure and that strong interactions block the spatial flexibility required for correct assembly and function.<\/jats:p>","DOI":"10.1128\/jb.01426-08","type":"journal-article","created":{"date-parts":[[2009,2,28]],"date-time":"2009-02-28T02:11:02Z","timestamp":1235787062000},"page":"2985-2992","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":21,"title":["Interactions between<i>Brucella suis<\/i>VirB8 and Its Homolog TraJ from the Plasmid pSB102 Underline the Dynamic Nature of Type IV Secretion Systems"],"prefix":"10.1128","volume":"191","author":[{"given":"Gise\u0300le","family":"Bourg","sequence":"first","affiliation":[{"name":"INSERM ESPRI 26, Universite\u0301 Montpellier 1 ERA 4204, Faculte\u0301 de Me\u0301decine, Avenue Kennedy, CS 83021, 30908 Ni\u0302mes Ce\u0301dex 2, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romain","family":"Sube","sequence":"additional","affiliation":[{"name":"INSERM ESPRI 26, Universite\u0301 Montpellier 1 ERA 4204, Faculte\u0301 de Me\u0301decine, Avenue Kennedy, CS 83021, 30908 Ni\u0302mes Ce\u0301dex 2, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"O'Callaghan","sequence":"additional","affiliation":[{"name":"INSERM ESPRI 26, Universite\u0301 Montpellier 1 ERA 4204, Faculte\u0301 de Me\u0301decine, Avenue Kennedy, CS 83021, 30908 Ni\u0302mes Ce\u0301dex 2, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilles","family":"Patey","sequence":"additional","affiliation":[{"name":"INSERM ESPRI 26, Universite\u0301 Montpellier 1 ERA 4204, Faculte\u0301 de Me\u0301decine, Avenue Kennedy, CS 83021, 30908 Ni\u0302mes Ce\u0301dex 2, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1038\/sj.emboj.7601983","volume":"27","year":"2008","unstructured":"Alix, E., and A. B. Blanc-Potard. 2008. Peptide-assisted degradation of the Salmonella MgtC virulence factor. EMBO J. 27 : 546-557.","journal-title":"EMBO J."},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00060-06"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.7.4255-4263.2000"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2008.07.001"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"2582","DOI":"10.1073\/pnas.0511216103","volume":"103","year":"2006","unstructured":"Bailey, S., D. Ward, R. Middleton, J. G. Grossmann, and P. C. Zambryski. 2006. Agrobacterium tumefaciens VirB8 structure reveals potential protein-protein interaction sites. Proc. Natl. Acad. Sci. USA 103 : 2582-2587.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_7_2","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1139\/o06-148","volume":"84","year":"2006","unstructured":"Baron, C. 2006. VirB8: a conserved type IV secretion system assembly factor and drug target. Biochem. Cell Biol. 84 : 890-899.","journal-title":"Biochem. Cell Biol."},{"key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/S1369-5274(00)00165-X","volume":"4","year":"2001","unstructured":"Boschiroli, M. L., V. Foulongne, and D. O'Callaghan. 2001. Brucellosis: a world wide zoonosis. Curr. Opin. Microbiol. 4 : 58-64.","journal-title":"Curr. Opin. Microbiol."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"1544","DOI":"10.1073\/pnas.032514299","volume":"99","year":"2002","unstructured":"Boschiroli, M. L., S. Ouahrani-Bettache, V. Foulongne, S. Michaux-Charachon, G. Bourg, A. lardet-Servent, C. Cazevieille, J. P. Liautard, M. Ramuz, and D. O'Callaghan. 2002. The Brucella suis virB operon is induced intracellularly in macrophages. Proc. Natl. Acad. Sci. USA 99 : 1544-1549.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.74.1.108-117.2006"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro753"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1095211"},{"key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"17228","DOI":"10.1073\/pnas.0405843101","volume":"101","year":"2004","unstructured":"Cascales, E., and P. J. Christie. 2004. Agrobacterium VirB10, an ATP energy sensor required for type IV secretion. Proc. Natl. Acad. Sci. USA 101 : 17228-17233.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.58.030603.123630"},{"key":"e_1_3_2_15_2","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1046\/j.1462-5822.2001.00102.x","volume":"3","year":"2001","unstructured":"Comerci, D. J., M. J. Martinez-Lorenzo, R. Sieira, J. P. Gorvel, and R. A. Ugalde. 2001. Essential role of the VirB machinery in the maturation of the Brucella abortus-containing vacuole. Cell. Microbiol. 3 : 159-168.","journal-title":"Cell. Microbiol."},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.3.758-763.2000"},{"key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0378-1097(03)00963-7","volume":"231","year":"2004","unstructured":"Delrue, R. M., P. Lestrate, A. Tibor, J. J. Letesson, and X. De Bolle. 2004. Brucella pathogenesis, genes identified from random large-scale screens. FEMS Microbiol. Lett. 231 : 1-12.","journal-title":"FEMS Microbiol. Lett."},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.20.5572-5582.2002"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.62.10.4135-4139.1994"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.3.1297-1303.2000"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1166101"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.11.5485-5493.1998"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.7.4102-4107.2000"},{"key":"e_1_3_2_24_2","first-page":"513","year":"1997","unstructured":"Hoover, D., and A. Friedlander. 1997. Brucellosis, p. 513-521. In F. Sidell, E. Takafuji, and D. Franz (ed.), Medical aspects of chemical and biological warfare. Office of the Surgeon General, Washington, DC.","journal-title":"Medical aspects of chemical and biological warfare"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.28326-0"},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"3148","DOI":"10.1105\/tpc.104.026476","volume":"16","year":"2004","unstructured":"Hwang, H. H., and S. B. Gelvin. 2004. Plant proteins that interact with VirB2, the Agrobacterium tumefaciens pilin protein, mediate plant transformation. Plant Cell 16 : 3148-3167.","journal-title":"Plant Cell"},{"key":"e_1_3_2_27_2","doi-asserted-by":"crossref","first-page":"11498","DOI":"10.1073\/pnas.0505290102","volume":"102","year":"2005","unstructured":"Judd, P. K., R. B. Kumar, and A. Das. 2005. Spatial location and requirements for the assembly of the Agrobacterium tumefaciens type IV secretion apparatus. Proc. Natl. Acad. Sci. USA 102 : 11498-11503.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","first-page":"5752","DOI":"10.1073\/pnas.95.10.5752","volume":"95","year":"1998","unstructured":"Karimova, G., J. Pidoux, A. Ullmann, and D. Ladant. 1998. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc. Natl. Acad. Sci. USA 95 : 5752-5756.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.7.2233-2243.2005"},{"key":"e_1_3_2_30_2","doi-asserted-by":"crossref","first-page":"15711","DOI":"10.1073\/pnas.232454299","volume":"99","year":"2002","unstructured":"K\u00f6hler, S., V. Foulongne, S. Ouarhani-Bettache, G. Bourg, J. Teyssier, M. Ramuz, and J.-P. Liautard. 2002. The analysis of the intramacrophagic virulome of Brucella suis deciphers the environment encountered by the pathogen inside the macrophage host cell. Proc. Natl. Acad. Sci. USA 99 : 15711-15716.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_31_2","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1046\/j.1365-2958.2000.01876.x","volume":"36","year":"2000","unstructured":"Kumar, R. B., Y. H. Xie, and A. Das. 2000. Subcellular localization of the Agrobacterium tumefaciens T-DNA transport pore proteins: VirB8 is essential for the assembly of the transport pore. Mol. Microbiol. 36 : 608-617.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_32_2","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1046\/j.1365-2958.2000.02150.x","volume":"38","year":"2000","unstructured":"Lestrate, P., R. M. Delrue, I. Danese, C. Didembourg, B. Taminiau, P. Mertens, X. De Bolle, A. Tibor, C. M. Tang, and J. J. Letesson. 2000. Identification and characterization of in vivo attenuated mutants of Brucella melitensis. Mol. Microbiol. 38 : 543-551.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.11.3259-3269.2003"},{"key":"e_1_3_2_34_2","unstructured":"A short course in bacterial genetics: a laboratory manual and handbook for Escherichia coli and related bacteria. 1992"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.1.486-493.2001"},{"key":"e_1_3_2_36_2","doi-asserted-by":"crossref","first-page":"3685","DOI":"10.1016\/S0021-9258(18)97200-5","volume":"240","year":"1965","unstructured":"Neu, H. C., and L. A. Heppel. 1965. The release of enzymes of E. coli by osmotic shock during the formation of spheroplasts. J. Biol. Chem. 240 : 3685-3692.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_37_2","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1046\/j.1365-2958.1999.01569.x","volume":"33","year":"1999","unstructured":"O'Callaghan, D., C. Cazevieille, A. Allardet-Servent, M. L. Boschiroli, G. Bourg, V. Foulongne, P. Frutos, Y. Kulakov, and M. Ramuz. 1999. A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis. Mol. Microbiol. 33 : 1210-1220.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_38_2","doi-asserted-by":"crossref","first-page":"7252","DOI":"10.1073\/pnas.0600862103","volume":"103","year":"2006","unstructured":"Paschos, A., G. Patey, D. Sivanesan, C. Gao, R. Bayliss, G. Waksman, D. O'Callaghan, and C. Baron. 2006. Dimerization and interactions of Brucella suis VirB8 with VirB4 and VirB10 are required for its biological activity. Proc. Natl. Acad. Sci. USA 103 : 7252-7257.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00584-06"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.5.2387-2392.1998"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.12.5711-5724.1998"},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.8.4041-4047.1999"},{"key":"e_1_3_2_43_2","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1046\/j.1365-2958.2003.03549.x","volume":"49","year":"2003","unstructured":"Rohde, M., J. P\u00fcls, R. Buhrdorf, W. Fischer, and R. Haas. 2003. A novel sheathed surface organelle of the Helicobacter pylori cag type IV secretion system. Mol. Microbiol. 49 : 219-234.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_44_2","doi-asserted-by":"crossref","first-page":"5169","DOI":"10.1093\/nar\/29.24.5169","volume":"29","year":"2001","unstructured":"Schneiker, S., M. Keller, M. Droge, E. Lanka, A. Puhler, and W. Selbitschka. 2001. The genetic organization and evolution of the broad host range mercury resistance plasmid pSB102 isolated from a microbial population residing in the rhizosphere of alfalfa. Nucleic Acids Res. 29 : 5169-5181.","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.17.4849-4855.2000"},{"key":"e_1_3_2_46_2","doi-asserted-by":"crossref","first-page":"1637","DOI":"10.1099\/00221287-148-6-1637","volume":"148","year":"2002","unstructured":"Tauch, A., S. Schneike, W. Selbitschka, A. Puhler, L. S. van Overbeek, K. Smalla, C. M. Thomas, M. J. Bailey, L. J. Forney, A. Weightman, P. Ceglowski, T. Pembroke, E. Tietze, G. Schroder, E. Lanka, and van J. D. Elsas. 2002. The complete nucleotide sequence and environmental distribution of the cryptic, conjugative, broad-host-range plasmid pIPO2 isolated from bacteria of the wheat rhizosphere. Microbiology 148 : 1637-1653.","journal-title":"Microbiology"},{"key":"e_1_3_2_47_2","doi-asserted-by":"crossref","first-page":"4596","DOI":"10.1073\/pnas.0408927102","volume":"102","year":"2005","unstructured":"Terradot, L., R. Bayliss, C. Oomen, G. A. Leonard, C. Baron, and G. Waksman. 2005. Structures of two core subunits of the bacterial type IV secretion system, VirB8 from Brucella suis and ComB10 from Helicobacter pylori. Proc. Natl. Acad. Sci. USA 102 : 4596-4601.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_48_2","doi-asserted-by":"crossref","first-page":"11493","DOI":"10.1073\/pnas.172390299","volume":"99","year":"2002","unstructured":"Ward, D. V., O. Draper, J. R. Zupan, and P. C. Zambryski. 2002. Peptide linkage mapping of the Agrobacterium tumefaciens vir-encoded type IV secretion system reveals protein subassemblies. Proc. Natl. Acad. Sci. USA 99 : 11493-11500.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_49_2","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1093\/clinids\/21.2.283","volume":"21","year":"1995","unstructured":"Young, E. J. 1995. An overview of human brucellosis. Clin. Infect. Dis. 21 : 283-289.","journal-title":"Clin. Infect. Dis."},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M502347200"}],"container-title":["Journal of Bacteriology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.01426-08","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.01426-08","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,11]],"date-time":"2024-03-11T00:00:45Z","timestamp":1710115245000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JB.01426-08"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,5]]},"references-count":49,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2009,5]]}},"alternative-id":["10.1128\/JB.01426-08"],"URL":"https:\/\/doi.org\/10.1128\/jb.01426-08","relation":{},"ISSN":["0021-9193","1098-5530"],"issn-type":[{"value":"0021-9193","type":"print"},{"value":"1098-5530","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,5]]},"assertion":[{"value":"2008-10-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2009-02-06","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-12-20","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}