{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T20:11:26Z","timestamp":1790799086479,"version":"4.1.0"},"reference-count":34,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,5,26]],"date-time":"2017-05-26T00:00:00Z","timestamp":1495756800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,5,26]],"date-time":"2017-05-26T00:00:00Z","timestamp":1495756800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Streptococcus pneumoniae<\/jats:italic> (the pneumococcus) is a human pathogen, accounting for massive global morbidity and mortality. Although asymptomatic colonization of the nasopharynx almost invariably precedes disease, the critical determinants enabling pneumococcal progression from this niche to cause invasive disease are poorly understood. One mechanism proposed to be central to this transition involves opacity phase variation, whereby pneumococci harvested from the nasopharynx are typically transparent, while those simultaneously harvested from the blood are opaque. Here, we used two dimensional-differential gel electrophoresis (2D-DIGE) to compare protein expression profiles of transparent and opaque variants of 3 pneumococcal strains, D39 (serotype 2), WCH43 (serotype 4) and WCH16 (serotype 6A) <jats:italic>in vitro<\/jats:italic>. One spot comprising a mixture of capsular polysaccharide biosynthesis protein and other proteins was significantly up-regulated in the opaque phenotype in all 3 strains; other proteins were differentially regulated in a strain-specific manner. We conclude that pneumococcal phase variation is a complex and multifactorial process leading to strain-specific pathogenicity.<\/jats:p>","DOI":"10.1038\/s41598-017-02465-x","type":"journal-article","created":{"date-parts":[[2017,5,22]],"date-time":"2017-05-22T14:37:39Z","timestamp":1495463859000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Proteomic comparisons of opaque and transparent variants of Streptococcus pneumoniae by two dimensional-differential gel electrophoresis"],"prefix":"10.1038","volume":"7","author":[{"given":"Melissa H.","family":"Chai","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Florian","family":"Weiland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard M.","family":"Harvey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Hoffmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abiodun D.","family":"Ogunniyi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James C.","family":"Paton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,5,26]]},"reference":[{"key":"2465_CR1","unstructured":"WHO. World Health Organization Media Centre; Pneumonia: Fact sheet No. 331 (2013)."},{"key":"2465_CR2","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1016\/S1473-3099(04)00938-7","volume":"4","author":"D Bogaert","year":"2004","unstructured":"Bogaert, D., De Groot, R. & Hermans, P. W. Streptococcus pneumoniae colonisation: the key to pneumococcal disease. Lancet Infect Dis \n                           4, 144\u2013154, doi:10.1016\/S1473-3099(04)00938-7 (2004).","journal-title":"Lancet Infect Dis"},{"key":"2465_CR3","doi-asserted-by":"crossref","first-page":"2582","DOI":"10.1128\/iai.62.6.2582-2589.1994","volume":"62","author":"JN Weiser","year":"1994","unstructured":"Weiser, J. N., Austrian, R., Sreenivasan, P. K. & Masure, H. R. Phase variation in pneumococcal opacity: relationship between colonial morphology and nasopharyngeal colonization. Infect Immun \n                           62, 2582\u20132589 (1994).","journal-title":"Infect Immun"},{"key":"2465_CR4","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1086\/514205","volume":"177","author":"JO Kim","year":"1998","unstructured":"Kim, J. O. & Weiser, J. N. Association of intrastrain phase variation in quantity of capsular polysaccharide and teichoic acid with the virulence of Streptococcus pneumoniae. J Infect Dis \n                           177, 368\u2013377, doi:10.1086\/jid.1998.177.issue-2 (1998).","journal-title":"J Infect Dis"},{"key":"2465_CR5","doi-asserted-by":"publisher","first-page":"6945","DOI":"10.1128\/IAI.73.10.6945-6951.2005","volume":"73","author":"DE Briles","year":"2005","unstructured":"Briles, D. E., Novak, L., Hotomi, M., van Ginkel, F. W. & King, J. Nasal colonization with Streptococcus pneumoniae includes subpopulations of surface and invasive pneumococci. Infect Immun \n                           73, 6945\u20136951, doi:10.1128\/IAI.73.10.6945-6951.2005 (2005).","journal-title":"Infect Immun"},{"key":"2465_CR6","doi-asserted-by":"publisher","first-page":"5055","DOI":"10.1038\/ncomms6055","volume":"5","author":"AS Manso","year":"2014","unstructured":"Manso, A. S. et al. A random six-phase switch regulates pneumococcal virulence via global epigenetic changes. Nat Commun \n                           5, 5055, doi:10.1038\/ncomms6055 (2014).","journal-title":"Nat Commun"},{"key":"2465_CR7","doi-asserted-by":"publisher","first-page":"4604","DOI":"10.1128\/IAI.68.8.4604-4610.2000","volume":"68","author":"K Overweg","year":"2000","unstructured":"Overweg, K. et al. Differential protein expression in phenotypic variants of Streptococcus pneumoniae. Infect Immun \n                           68, 4604\u20134610, doi:10.1128\/IAI.68.8.4604-4610.2000 (2000).","journal-title":"Infect Immun"},{"key":"2465_CR8","doi-asserted-by":"publisher","first-page":"5690","DOI":"10.1128\/IAI.68.10.5690-5695.2000","volume":"68","author":"KK Gosink","year":"2000","unstructured":"Gosink, K. K., Mann, E. R., Guglielmo, C., Tuomanen, E. I. & Masure, H. R. Role of novel choline binding proteins in virulence of Streptococcus pneumoniae. Infect Immun \n                           68, 5690\u20135695, doi:10.1128\/IAI.68.10.5690-5695.2000 (2000).","journal-title":"Infect Immun"},{"key":"2465_CR9","doi-asserted-by":"publisher","first-page":"14363","DOI":"10.1073\/pnas.2235844100","volume":"100","author":"FW van Ginkel","year":"2003","unstructured":"van Ginkel, F. W. et al. Pneumococcal carriage results in ganglioside-mediated olfactory tissue infection. 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PLoS One \n                           8, e68277, doi:10.1371\/journal.pone.0068277 (2013).","journal-title":"PLoS One"},{"key":"2465_CR12","doi-asserted-by":"publisher","first-page":"e70862","DOI":"10.1371\/journal.pone.0070862","volume":"8","author":"LK Mahdi","year":"2013","unstructured":"Mahdi, L. K., Ebrahimie, E., Adelson, D. L., Paton, J. C. & Ogunniyi, A. D. A transcription factor contributes to pathogenesis and virulence in Streptococcus pneumoniae. PLoS One \n                           8, e70862, doi:10.1371\/journal.pone.0070862 (2013).","journal-title":"PLoS One"},{"key":"2465_CR13","doi-asserted-by":"publisher","first-page":"646","DOI":"10.1128\/IAI.01161-07","volume":"76","author":"LK Mahdi","year":"2008","unstructured":"Mahdi, L. K., Ogunniyi, A. D., LeMessurier, K. S. & Paton, J. C. Pneumococcal virulence gene expression and host cytokine profiles during pathogenesis of invasive disease. 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Variable opacity (Opa) outer membrane proteins account for the cell tropisms displayed by Neisseria gonorrhoeae for human leukocytes and epithelial cells. Embo J \n                           12, 641\u2013650 (1993).","journal-title":"Embo J"},{"key":"2465_CR18","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1002\/j.1460-2075.1991.tb07649.x","volume":"10","author":"S Makino","year":"1991","unstructured":"Makino, S., van Putten, J. P. & Meyer, T. F. Phase variation of the opacity outer membrane protein controls invasion by Neisseria gonorrhoeae into human epithelial cells. Embo J \n                           10, 1307\u20131315 (1991).","journal-title":"Embo J"},{"key":"2465_CR19","doi-asserted-by":"publisher","first-page":"672","DOI":"10.1093\/infdis\/168.3.672","volume":"168","author":"JN Weiser","year":"1993","unstructured":"Weiser, J. N. Relationship between colony morphology and the life cycle of Haemophilus influenzae: the contribution of lipopolysaccharide phase variation to pathogenesis. J Infect Dis \n                           168, 672\u2013680, doi:10.1093\/infdis\/168.3.672 (1993).","journal-title":"J Infect Dis"},{"key":"2465_CR20","doi-asserted-by":"publisher","first-page":"5430","DOI":"10.1128\/IAI.69.9.5430-5439.2001","volume":"69","author":"JN Weiser","year":"2001","unstructured":"Weiser, J. N. et al. Changes in availability of oxygen accentuate differences in capsular polysaccharide expression by phenotypic variants and clinical isolates of Streptococcus pneumoniae. Infect Immun \n                           69, 5430\u20135439, doi:10.1128\/IAI.69.9.5430-5439.2001 (2001).","journal-title":"Infect Immun"},{"key":"2465_CR21","doi-asserted-by":"publisher","first-page":"6330","DOI":"10.1128\/JB.00707-08","volume":"190","author":"M Allegrucci","year":"2008","unstructured":"Allegrucci, M. & Sauer, K. Formation of Streptococcus pneumoniae non-phase-variable colony variants is due to increased mutation frequency present under biofilm growth conditions. J Bacteriol \n                           190, 6330\u20136339, doi:10.1128\/JB.00707-08 (2008).","journal-title":"J Bacteriol"},{"key":"2465_CR22","doi-asserted-by":"publisher","first-page":"8164","DOI":"10.1128\/JB.186.24.8164-8171.2004","volume":"186","author":"AE Belanger","year":"2004","unstructured":"Belanger, A. E., Clague, M. J., Glass, J. I. & Leblanc, D. J. Pyruvate oxidase is a determinant of Avery\u2019s rough morphology. J Bacteriol \n                           186, 8164\u20138171, doi:10.1128\/JB.186.24.8164-8171.2004 (2004).","journal-title":"J Bacteriol"},{"key":"2465_CR23","doi-asserted-by":"publisher","first-page":"6532","DOI":"10.1128\/JB.00813-07","volume":"189","author":"G Regev-Yochay","year":"2007","unstructured":"Regev-Yochay, G., Trzcinski, K., Thompson, C. M., Lipsitch, M. & Malley, R. SpxB is a suicide gene of Streptococcus pneumoniae and confers a selective advantage in an in vivo competitive colonization model. J Bacteriol \n                           189, 6532\u20136539, doi:10.1128\/JB.00813-07 (2007).","journal-title":"J Bacteriol"},{"key":"2465_CR24","first-page":"703","volume":"39","author":"SM Bae","year":"2006","unstructured":"Bae, S. M., Yeon, S. M., Kim, T. S. & Lee, K. J. The effect of protein expression of Streptococcus pneumoniae by blood. J Biochem Mol Biol \n                           39, 703\u2013708 (2006).","journal-title":"J Biochem Mol Biol"},{"key":"2465_CR25","doi-asserted-by":"publisher","first-page":"4392","DOI":"10.1128\/JB.184.16.4392-4399.2002","volume":"184","author":"CD Pericone","year":"2002","unstructured":"Pericone, C. D., Bae, D., Shchepetov, M., McCool, T. & Weiser, J. N. Short-sequence tandem and nontandem DNA repeats and endogenous hydrogen peroxide production contribute to genetic instability of Streptococcus pneumoniae. J Bacteriol \n                           184, 4392\u20134399, doi:10.1128\/JB.184.16.4392-4399.2002 (2002).","journal-title":"J Bacteriol"},{"key":"2465_CR26","doi-asserted-by":"publisher","first-page":"6390","DOI":"10.1128\/JB.00995-10","volume":"192","author":"S Ramos-Montanez","year":"2010","unstructured":"Ramos-Montanez, S., Kazmierczak, K. M., Hentchel, K. L. & Winkler, M. E. Instability of ackA (acetate kinase) mutations and their effects on acetyl phosphate and ATP amounts in Streptococcus pneumoniae D39. J Bacteriol \n                           192, 6390\u20136400, doi:10.1128\/JB.00995-10 (2010).","journal-title":"J Bacteriol"},{"key":"2465_CR27","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1111\/j.1365-2958.2007.06082.x","volume":"67","author":"S Ramos-Montanez","year":"2008","unstructured":"Ramos-Montanez, S. et al. Polymorphism and regulation of the spxB (pyruvate oxidase) virulence factor gene by a CBS-HotDog domain protein (SpxR) in serotype 2 Streptococcus pneumoniae. Mol Microbiol \n                           67, 729\u2013746, doi:10.1111\/mmi.2008.67.issue-4 (2008).","journal-title":"Mol Microbiol"},{"key":"2465_CR28","doi-asserted-by":"publisher","first-page":"02005","DOI":"10.1128\/IAI.02005-14","volume":"2014","author":"P Gaspar","year":"2014","unstructured":"Gaspar, P., Al-Bayati, F. A., Andrew, P. W., Neves, A. R. & Yesilkaya, H. Lactate dehydrogenase is the key enzyme for pneumococcal pyruvate metabolism and pneumococcal survival in blood. Infect Immun \n                           2014, 02005\u201302014, doi:10.1128\/IAI.02005-14 (2014).","journal-title":"Infect Immun"},{"key":"2465_CR29","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1128\/IAI.68.1.133-140.2000","volume":"68","author":"AM Berry","year":"2000","unstructured":"Berry, A. M. & Paton, J. C. Additive attenuation of virulence of Streptococcus pneumoniae by mutation of the genes encoding pneumolysin and other putative pneumococcal virulence proteins. Infect Immun \n                           68, 133\u2013140, doi:10.1128\/IAI.68.1.133-140.2000 (2000).","journal-title":"Infect Immun"},{"key":"2465_CR30","doi-asserted-by":"publisher","first-page":"508","DOI":"10.1016\/0006-3002(60)90205-5","volume":"39","author":"S Lacks","year":"1960","unstructured":"Lacks, S. & Hotchkiss, R. D. A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta \n                           39, 508\u2013518, doi:10.1016\/0006-3002(60)90205-5 (1960).","journal-title":"Biochim Biophys Acta"},{"issue":"Suppl 1","key":"2465_CR31","doi-asserted-by":"publisher","first-page":"S122","DOI":"10.1002\/elps.200900051","volume":"30","author":"A Gorg","year":"2009","unstructured":"Gorg, A., Drews, O., Luck, C., Weiland, F. & Weiss, W. 2-DE with IPGs. Electrophoresis \n                           30(Suppl 1), S122\u2013132, doi:10.1002\/elps.200900051 (2009).","journal-title":"Electrophoresis"},{"key":"2465_CR32","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1186\/1471-2164-15-769","volume":"15","author":"LK Mahdi","year":"2014","unstructured":"Mahdi, L. K. et al. A functional genomics catalogue of activated transcription factors during pathogenesis of pneumococcal disease. BMC Genomics \n                           15, 769, doi:10.1186\/1471-2164-15-769 (2014).","journal-title":"BMC Genomics"},{"key":"2465_CR33","doi-asserted-by":"publisher","first-page":"538","DOI":"10.1128\/AAC.46.2.538-542.2002","volume":"46","author":"SJ Beard","year":"2002","unstructured":"Beard, S. J., Salisbury, V., Lewis, R. J., Sharpe, J. A. & MacGowan, A. P. Expression of lux genes in a clinical isolate of Streptococcus pneumoniae: using bioluminescence to monitor gemifloxacin activity. Antimicrob Agents Chemother \n                           46, 538\u2013542, doi:10.1128\/AAC.46.2.538-542.2002 (2002).","journal-title":"Antimicrob Agents Chemother"},{"key":"2465_CR34","doi-asserted-by":"publisher","first-page":"14031","DOI":"10.1074\/jbc.275.19.14031","volume":"275","author":"S Fillinger","year":"2000","unstructured":"Fillinger, S. et al. Two glyceraldehyde-3-phosphate dehydrogenases with opposite physiological roles in a nonphotosynthetic bacterium. J Biol Chem \n                           275, 14031\u201314037, doi:10.1074\/jbc.275.19.14031 (2000).","journal-title":"J Biol Chem"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-02465-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-02465-x","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-02465-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T04:41:57Z","timestamp":1671770517000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-02465-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,5,26]]},"references-count":34,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["2465"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-02465-x","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,5,26]]},"assertion":[{"value":"6 December 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 April 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 May 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"2453"}}