Infection of host tissues by Staphylococcus aureus and S. epidermidis requires an unusual family of staphylococcal adhesive proteins that contain long stretches of serine-aspartate dipeptide-repeats (SDR). The prototype member of this family is clumping factor A (ClfA), a key virulence factor that mediates adhesion to host tissues by binding to extracellular matrix proteins such as fibrinogen. However, the biological siginificance of the SDR-domain and its implication for pathogenesis remain poorly understood. Here, we identified two novel bacterial glycosyltransferases, SdgA and SdgB, which modify all SDR-proteins in these two bacterial species. Genetic and biochemical data demonstrated that these two glycosyltransferases directly bind and covalently link N-acetylglucosamine (GlcNAc) moieties to the SDR-domain in a step-wise manner, with SdgB appending the sugar residues proximal to the target Ser-Asp repeats, followed by additional modification by SdgA. GlcNAc-modification of SDR-proteins by SdgB creates an immunodominant epitope for highly opsonic human antibodies, which represent up to 1% of total human IgG. Deletion of these glycosyltransferases renders SDR-proteins vulnerable to proteolysis by human neutrophil-derived cathepsin G. Thus, SdgA and SdgB glycosylate staphylococcal SDR-proteins, which protects them against host proteolytic activity, and yet generates major eptopes for the human anti-staphylococcal antibody response, which may represent an ongoing competition between host and pathogen.
References
[1]
Chambers HF, DeLeo FR (2009) Waves of resistance: Staphylococcus aureus in the antibiotic era. Nature reviews Microbiology 7: 629–641. doi: 10.1038/nrmicro2200
[2]
Foster TJ (2009) Colonization and infection of the human host by staphylococci: adhesion, survival and immune evasion. Vet Dermatol 20: 456–470. doi: 10.1111/j.1365-3164.2009.00825.x
[3]
McAdow M, Kim HK, Dedent AC, Hendrickx AP, Schneewind O, et al. (2011) Preventing Staphylococcus aureus sepsis through the inhibition of its agglutination in blood. PLoS pathogens 7: e1002307. doi: 10.1371/journal.ppat.1002307
[4]
Foster TJ, Hook M (1998) Surface protein adhesins of Staphylococcus aureus. Trends Microbiol 6: 484–488. doi: 10.1016/s0966-842x(98)01400-0
[5]
Cheng AG, Kim HK, Burts ML, Krausz T, Schneewind O, et al. (2009) Genetic requirements for Staphylococcus aureus abscess formation and persistence in host tissues. FASEB journal: official publication of the Federation of American Societies for Experimental Biology 23: 3393–3404. doi: 10.1096/fj.09-135467
[6]
McCrea KW, Hartford O, Davis S, Eidhin DN, Lina G, et al. (2000) The serine-aspartate repeat (Sdr) protein family in Staphylococcus epidermidis. Microbiology 146 (Pt 7): 1535–1546.
[7]
Hartford O, Francois P, Vaudaux P, Foster TJ (1997) The dipeptide repeat region of the fibrinogen-binding protein (clumping factor) is required for functional expression of the fibrinogen-binding domain on the Staphylococcus aureus cell surface. Mol Microbiol 25: 1065–1076. doi: 10.1046/j.1365-2958.1997.5291896.x
[8]
Cooper JE, Feil EJ (2006) The phylogeny of Staphylococcus aureus - which genes make the best intra-species markers? Microbiology 152: 1297–1305. doi: 10.1099/mic.0.28620-0
[9]
Marraffini LA, DeDent AC, Schneewind O (2006) Sortases and the art of anchoring proteins to the envelopes of Gram-positive bacteria. Microbiol Mol Biol Rev 70: 192–221. doi: 10.1128/mmbr.70.1.192-221.2006
[10]
Josefsson E, McCrea KW, Ni Eidhin D, O′Connell D, Cox J, et al. (1998) Three new members of the serine-aspartate repeat protein multigene family of Staphylococcus aureus. Microbiology 144 (Pt 12): 3387–3395. doi: 10.1099/00221287-144-12-3387
[11]
Mazmanian SK, Liu G, Jensen ER, Lenoy E, Schneewind O (2000) Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proceedings of the National Academy of Sciences of the United States of America 97: 5510–5515. doi: 10.1073/pnas.080520697
[12]
Fitzgerald JR, Loughman A, Keane F, Brennan M, Knobel M, et al. (2006) Fibronectin-binding proteins of Staphylococcus aureus mediate activation of human platelets via fibrinogen and fibronectin bridges to integrin GPIIb/IIIa and IgG binding to the FcgammaRIIa receptor. Molecular microbiology 59: 212–230. doi: 10.1111/j.1365-2958.2005.04922.x
[13]
Brown S, Xia G, Luhachack LG, Campbell J, Meredith TC, et al. (2012) Methicillin resistance in Staphylococcus aureus requires glycosylated wall teichoic acids. Proceedings of the National Academy of Sciences of the United States of America 109: 18909–18914. doi: 10.1073/pnas.1209126109
[14]
Xia G, Maier L, Sanchez-Carballo P, Li M, Otto M, et al. (2010) Glycosylation of wall teichoic acid in Staphylococcus aureus by TarM. J Biol Chem 285: 13405–13415. doi: 10.1074/jbc.m109.096172
[15]
Shah SR (2008) A newer immunoglobulin intravenous (IGIV) - Gammagard liquid 10%: evaluation of efficacy, safety, tolerability and impact on patient care. Expert opinion on biological therapy 8: 799–804. doi: 10.1517/14712598.8.6.799
[16]
Schwefel D, Maierhofer C, Beck JG, Seeberger S, Diederichs K, et al. (2010) Structural basis of multivalent binding to wheat germ agglutinin. Journal of the American Chemical Society 132: 8704–8719. doi: 10.1021/ja101646k
[17]
Zhou M, Wu H (2009) Glycosylation and biogenesis of a family of serine-rich bacterial adhesins. Microbiology 155: 317–327. doi: 10.1099/mic.0.025221-0
[18]
Charbonneau ME, Mourez M (2008) The Escherichia coli AIDA-I autotransporter undergoes cytoplasmic glycosylation independently of export. Research in microbiology 159: 537–544. doi: 10.1016/j.resmic.2008.06.009
[19]
Lindenthal C, Elsinghorst EA (2001) Enterotoxigenic Escherichia coli TibA glycoprotein adheres to human intestine epithelial cells. Infection and immunity 69: 52–57. doi: 10.1128/iai.69.1.52-57.2001
[20]
Wu H, Mintz KP, Ladha M, Fives-Taylor PM (1998) Isolation and characterization of Fap1, a fimbriae-associated adhesin of Streptococcus parasanguis FW213. Mol Microbiol 28: 487–500. doi: 10.1046/j.1365-2958.1998.00805.x
[21]
Plummer C, Wu H, Kerrigan SW, Meade G, Cox D, et al. (2005) A serine-rich glycoprotein of Streptococcus sanguis mediates adhesion to platelets via GPIb. Br J Haematol 129: 101–109. doi: 10.1111/j.1365-2141.2005.05421.x
[22]
Bensing BA, Gibson BW, Sullam PM (2004) The Streptococcus gordonii platelet binding protein GspB undergoes glycosylation independently of export. J Bacteriol 186: 638–645. doi: 10.1128/jb.186.3.638-645.2004
[23]
Siboo IR, Chaffin DO, Rubens CE, Sullam PM (2008) Characterization of the accessory Sec system of Staphylococcus aureus. J Bacteriol 190: 6188–6196. doi: 10.1128/jb.00300-08
[24]
Zhou M, Zhu F, Dong S, Pritchard DG, Wu H (2010) A novel glucosyltransferase is required for glycosylation of a serine-rich adhesin and biofilm formation by Streptococcus parasanguinis. The Journal of biological chemistry 285: 12140–12148. doi: 10.1074/jbc.m109.066928
[25]
Sanchez CJ, Shivshankar P, Stol K, Trakhtenbroit S, Sullam PM, et al. (2010) The pneumococcal serine-rich repeat protein is an intra-species bacterial adhesin that promotes bacterial aggregation in vivo and in biofilms. PLoS pathogens 6: e1001044. doi: 10.1371/journal.ppat.1001044
[26]
Abraham NM, Jefferson KK (2012) Staphylococcus aureus clumping factor B mediates biofilm formation in the absence of calcium. Microbiology 158: 1504–1512. doi: 10.1099/mic.0.057018-0
[27]
Sharp JA, Echague CG, Hair PS, Ward MD, Nyalwidhe JO, et al. (2012) Staphylococcus aureus surface protein SdrE binds complement regulator factor H as an immune evasion tactic. PloS one 7: e38407. doi: 10.1371/journal.pone.0038407
[28]
Raymond WW, Trivedi NN, Makarova A, Ray M, Craik CS, et al. (2010) How immune peptidases change specificity: cathepsin G gained tryptic function but lost efficiency during primate evolution. Journal of immunology 185: 5360–5368. doi: 10.4049/jimmunol.1002292
[29]
Mandlik A, Livny J, Robins WP, Ritchie JM, Mekalanos JJ, et al. (2011) RNA-Seq-based monitoring of infection-linked changes in Vibrio cholerae gene expression. Cell Host Microbe 10: 165–174. doi: 10.1016/j.chom.2011.07.007
[30]
Pishchany G, McCoy AL, Torres VJ, Krause JC, Crowe JE Jr, et al. (2010) Specificity for human hemoglobin enhances Staphylococcus aureus infection. Cell host & microbe 8: 544–550. doi: 10.1016/j.chom.2010.11.002
[31]
Spaan AN, Henry T, van Rooijen WJ, Perret M, Badiou C, et al. (2013) The staphylococcal toxin panton-valentine leukocidin targets human c5a receptors. Cell host & microbe 13: 584–594. doi: 10.1016/j.chom.2013.04.006
[32]
Monk IR, Shah IM, Xu M, Tan MW, Foster TJ (2012) Transforming the untransformable: application of direct transformation to manipulate genetically Staphylococcus aureus and Staphylococcus epidermidis. mBio 3: e00277–11. doi: 10.1128/mbio.00277-11
[33]
Kwakkenbos MJ, Diehl SA, Yasuda E, Bakker AQ, van Geelen CM, et al. (2010) Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming. Nat Med 16: 123–128. doi: 10.1038/nm.2071
[34]
Meijer PJ, Andersen PS, Haahr Hansen M, Steinaa L, Jensen A, et al. (2006) Isolation of human antibody repertoires with preservation of the natural heavy and light chain pairing. Journal of molecular biology 358: 764–772. doi: 10.1016/j.jmb.2006.02.040
[35]
Meijer PJ, Nielsen LS, Lantto J, Jensen A (2009) Human antibody repertoires. Methods in molecular biology 525: : 261–277, xiv.
[36]
Phu L, Izrael-Tomasevic A, Matsumoto ML, Bustos D, Dynek JN, et al.. (2011) Improved quantitative mass spectrometry methods for characterizing complex ubiquitin signals. Molecular & cellular proteomics: MCP 10: M110 003756.
[37]
Nesvizhskii AI, Keller A, Kolker E, Aebersold R (2003) A statistical model for identifying proteins by tandem mass spectrometry. Analytical chemistry 75: 4646–4658. doi: 10.1021/ac0341261
[38]
Nakamura GR, Byrn R, Wilkes DM, Fox JA, Hobbs MR, et al. (1993) Strain specificity and binding affinity requirements of neutralizing monoclonal antibodies to the C4 domain of gp120 from human immunodeficiency virus type 1. J Virol 67: 6179–6191.
[39]
Tattevin P, Basuino L, Bauer D, Diep BA, Chambers HF (2010) Ceftobiprole is superior to vancomycin, daptomycin, and linezolid for treatment of experimental endocarditis in rabbits caused by methicillin-resistant Staphylococcus aureus. Antimicrobial agents and chemotherapy 54: 610–613. doi: 10.1128/aac.00886-09
[40]
McAleese FM, Walsh EJ, Sieprawska M, Potempa J, Foster TJ (2001) Loss of clumping factor B fibrinogen binding activity by Staphylococcus aureus involves cessation of transcription, shedding and cleavage by metalloprotease. The Journal of biological chemistry 276: 29969–29978. doi: 10.1074/jbc.m102389200
