{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T02:01:53Z","timestamp":1793325713954,"version":"4.5.0"},"reference-count":38,"publisher":"National Academy of Sciences","issue":"8","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2013,2,19]]},"abstract":"<jats:p>\n                    Natural transformation is a dominant force in bacterial evolution by promoting horizontal gene transfer. This process may have devastating consequences, such as the spread of antibiotic resistance or the emergence of highly virulent clones. However, uptake and recombination of foreign DNA are most often deleterious to competent species. Therefore, model naturally transformable Gram-negative bacteria, including the human pathogen\n                    <jats:italic>Neisseria meningitidis<\/jats:italic>\n                    , have evolved means to preferentially take up homotypic DNA containing short and genus-specific sequence motifs. Despite decades of intense investigations, the DNA uptake sequence receptor in\n                    <jats:italic>Neisseria<\/jats:italic>\n                    species has remained elusive. We show here, using a multidisciplinary approach combining biochemistry, molecular genetics, and structural biology, that meningococcal type IV pili bind DNA through the minor pilin ComP via an electropositive stripe that is predicted to be exposed on the filaments surface and that ComP displays an exquisite binding preference for DNA uptake sequence. Our findings illuminate the earliest step in natural transformation, reveal an unconventional mechanism for DNA binding, and suggest that selective DNA uptake is more widespread than previously thought.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1218832110","type":"journal-article","created":{"date-parts":[[2013,2,6]],"date-time":"2013-02-06T01:48:25Z","timestamp":1360115305000},"page":"3065-3070","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":200,"title":["Specific DNA recognition mediated by a type IV pilin"],"prefix":"10.1073","volume":"110","author":[{"given":"Ana","family":"Cehovin","sequence":"first","affiliation":[{"name":"Medical Research Council Centre for Molecular Bacteriology and Infection, Section of Microbiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter J.","family":"Simpson","sequence":"additional","affiliation":[{"name":"Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, United Kingdom; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melanie A.","family":"McDowell","sequence":"additional","affiliation":[{"name":"Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel R.","family":"Brown","sequence":"additional","affiliation":[{"name":"Medical Research Council Centre for Molecular Bacteriology and Infection, Section of Microbiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rossella","family":"Noschese","sequence":"additional","affiliation":[{"name":"Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, United Kingdom; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitchell","family":"Pallett","sequence":"additional","affiliation":[{"name":"Medical Research Council Centre for Molecular Bacteriology and Infection, Section of Microbiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob","family":"Brady","sequence":"additional","affiliation":[{"name":"Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, United Kingdom; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geoffrey S.","family":"Baldwin","sequence":"additional","affiliation":[{"name":"Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, United Kingdom; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan M.","family":"Lea","sequence":"additional","affiliation":[{"name":"Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen J.","family":"Matthews","sequence":"additional","affiliation":[{"name":"Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, United Kingdom; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladimir","family":"Pelicic","sequence":"additional","affiliation":[{"name":"Medical Research Council Centre for Molecular Bacteriology and Infection, Section of Microbiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2013,2,5]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1016\/j.resmic.2007.09.004","article-title":"Natural genetic transformation: Prevalence, mechanisms and function","volume":"158","author":"Johnsborg O","year":"2007","unstructured":"O Johnsborg, V Eldholm, LS H\u00e5varstein, Natural genetic transformation: Prevalence, mechanisms and function. 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Infect Immun 78, 3053\u20133063 (2010).","journal-title":"Infect Immun"},{"key":"e_1_3_4_22_2","doi-asserted-by":"crossref","first-page":"15276","DOI":"10.1073\/pnas.261574998","article-title":"Neisseria gonorrhoeae PilV, a type IV pilus-associated protein essential to human epithelial cell adherence","volume":"98","author":"Winther-Larsen HC","year":"2001","unstructured":"HC Winther-Larsen, et al., Neisseria gonorrhoeae PilV, a type IV pilus-associated protein essential to human epithelial cell adherence. 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PLoS One 5, e11835 (2010).","journal-title":"PLoS One"},{"key":"e_1_3_4_27_2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1038\/nrmicro885","article-title":"Type IV pilus structure and bacterial pathogenicity","volume":"2","author":"Craig L","year":"2004","unstructured":"L Craig, ME Pique, JA Tainer, Type IV pilus structure and bacterial pathogenicity. Nat Rev Microbiol 2, 363\u2013378 (2004).","journal-title":"Nat Rev Microbiol"},{"key":"e_1_3_4_28_2","first-page":"13","article-title":"Analysis of protein-DNA interactions using surface plasmon resonance","volume":"104","author":"Majka J","year":"2007","unstructured":"J Majka, C Speck, Analysis of protein-DNA interactions using surface plasmon resonance. Adv Biochem Eng Biotechnol 104, 13\u201336 (2007).","journal-title":"Adv Biochem Eng Biotechnol"},{"key":"e_1_3_4_29_2","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.molcel.2006.07.004","article-title":"Type IV pilus structure by cryo-electron microscopy and crystallography: Implications for pilus assembly and functions","volume":"23","author":"Craig L","year":"2006","unstructured":"L Craig, et al., Type IV pilus structure by cryo-electron microscopy and crystallography: Implications for pilus assembly and functions. Mol Cell 23, 651\u2013662 (2006).","journal-title":"Mol Cell"},{"key":"e_1_3_4_30_2","doi-asserted-by":"crossref","first-page":"e39742","DOI":"10.1371\/journal.pone.0039742","article-title":"Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis","volume":"7","author":"Ambur OH","year":"2012","unstructured":"OH Ambur, SA Frye, M Nilsen, E Hovland, T T\u00f8njum, Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis. PLoS One 7, e39742 (2012).","journal-title":"PLoS One"},{"key":"e_1_3_4_31_2","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1111\/j.1365-2958.2005.04838.x","article-title":"The frequency and rate of pilin antigenic variation in Neisseria gonorrhoeae","volume":"58","author":"Criss AK","year":"2005","unstructured":"AK Criss, KA Kline, HS Seifert, The frequency and rate of pilin antigenic variation in Neisseria gonorrhoeae. Mol Microbiol 58, 510\u2013519 (2005).","journal-title":"Mol Microbiol"},{"key":"e_1_3_4_32_2","first-page":"389","article-title":"Identification of Eikenella corrodens and Cardiobacterium hominis by genetic transformation","volume":"93","author":"T\u00f8njum T","year":"1985","unstructured":"T T\u00f8njum, N Hagen, K B\u00f8vre, Identification of Eikenella corrodens and Cardiobacterium hominis by genetic transformation. Acta Pathol Microbiol Immunol Scand [B] 93, 389\u2013394 (1985).","journal-title":"Acta Pathol Microbiol Immunol Scand [B]"},{"key":"e_1_3_4_33_2","doi-asserted-by":"crossref","first-page":"4993","DOI":"10.1128\/iai.64.12.4993-4999.1996","article-title":"Identification of four complete type 4 pilin genes in a single Kingella denitrificans genome","volume":"64","author":"Weir S","year":"1996","unstructured":"S Weir, LW Lee, CF Marrs, Identification of four complete type 4 pilin genes in a single Kingella denitrificans genome. Infect Immun 64, 4993\u20134999 (1996).","journal-title":"Infect Immun"},{"key":"e_1_3_4_34_2","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1016\/j.cell.2010.11.035","article-title":"Meningococcus Hijacks a \u03b22-adrenoceptor\/\u03b2-Arrestin pathway to cross brain microvasculature endothelium","volume":"143","author":"Coureuil M","year":"2010","unstructured":"M Coureuil, et al., Meningococcus Hijacks a \u03b22-adrenoceptor\/\u03b2-Arrestin pathway to cross brain microvasculature endothelium. Cell 143, 1149\u20131160 (2010).","journal-title":"Cell"},{"key":"e_1_3_4_35_2","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1101\/gr.664303","article-title":"Large-scale analysis of the meningococcus genome by gene disruption: Resistance to complement-mediated lysis","volume":"13","author":"Geoffroy MC","year":"2003","unstructured":"MC Geoffroy, S Floquet, A M\u00e9tais, X Nassif, V Pelicic, Large-scale analysis of the meningococcus genome by gene disruption: Resistance to complement-mediated lysis. 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