1887

Microbiology Society logo

Volume 168, Issue 5

The serotype a-EmaA adhesin of does not require O-PS synthesis for collagen binding activity Open Access

Abstract

, a causative agent of periodontitis and non-oral diseases, synthesizes a trimeric extracellular matrix protein adhesin A (EmaA) that mediates collagen binding and biofilm formation. EmaA is found as two molecular forms, which correlate with the serotype of the bacterium. The canonical protein (b-EmaA), associated with serotypes b and c, has a monomeric molecular mass of 202 kDa. The collagen binding activity of b-EmaA is dependent on the presence of O-polysaccharide (O-PS), whereas biofilm activity is independent of O-PS synthesis. The EmaA associated with serotype a strains (a-EmaA) has a monomeric molecular mass of 173 kDa and differs in the amino acid sequence of the functional domain of the protein. In this study, a- was confirmed to encode a protein that forms antenna-like appendages on the surface of the bacterium, which were found to be important for both collagen binding and biofilm formation. In an O-PS-deficient talose biosynthetic () mutant strain, the electrophoretic mobility of the a-EmaA monomers was altered and the amount of membrane-associated EmaA was decreased when compared to the parent strain. The mass of biofilm formed remained unchanged. Interestingly, the collagen binding activity of the mutant strain was similar to the activity associated with the parent strain, which differs from that observed with the canonical b-EmaA isoform. These data suggest that the properties of the a-EmaA isoform are like those of b-EmaA, with the exception that collagen binding activity is independent of the presence or absence of the O-PS.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): adhesin , collagen , endocarditis , glycosylation , Gram-negative bacterium , O-polysaccharides and periodontal disease
Funding
This study was supported by the:
  • National Institute of Dental and Craniofacial Research (Award R01-DE024554)
    • Principle Award Recipient: KeithP. Mintz
© 2022 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.001191
2022-05-12
2024-05-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/168/5/mic001191.html?itemId=/content/journal/micro/10.1099/mic.0.001191&mimeType=html&fmt=ahah

References

  1. Lamont RJ, Koo H, Hajishengallis G. The oral microbiota: dynamic communities and host interactions. Nat Rev Microbiol 2018; 16:745–759 [View Article] [PubMed]
     [Google Scholar]
  2. Hajishengallis G. Immunomicrobial pathogenesis of periodontitis: keystones, pathobionts, and host response. Trends Immunol 2014; 35:3–11 [View Article] [PubMed]
     [Google Scholar]
  3. Sen Yew H, Chambers ST, Roberts SA, Holland DJ, Julian KA et al. Association between HACEK bacteraemia and endocarditis. J Med Microbiol 2014; 63:892–895 [View Article] [PubMed]
     [Google Scholar]
  4. Tang G, Kitten T, Munro CL, Wellman GC, Mintz KP. EmaA, a potential virulence determinant of Aggregatibacter actinomycetemcomitans in infective endocarditis. Infect Immun 2008; 76:2316–2324 [View Article] [PubMed]
     [Google Scholar]
  5. Mintz KP. Identification of an extracellular matrix protein adhesin, EmaA, which mediates the adhesion of Actinobacillus actinomycetemcomitans to collagen. Microbiology (Reading) 2004; 150:2677–2688 [View Article] [PubMed]
     [Google Scholar]
  6. Ruiz T, Lenox C, Radermacher M, Mintz KP. Novel surface structures are associated with the adhesion of Actinobacillus actinomycetemcomitans to collagen. Infect Immun 2006; 74:6163–6170 [View Article] [PubMed]
     [Google Scholar]
  7. Danforth DR, Tang-Siegel G, Ruiz T, Mintz KP. A nonfimbrial adhesin of Aggregatibacter actinomycetemcomitans mediates biofilm biogenesis. Infect Immun 2019; 87:e00704-18 [View Article] [PubMed]
     [Google Scholar]
  8. Hoiczyk E, Roggenkamp A, Reichenbecher M, Lupas A, Heesemann J. Structure and sequence analysis of Yersinia YadA and Moraxella UspAs reveal a novel class of adhesins. EMBO J 2000; 19:5989–5999 [View Article] [PubMed]
     [Google Scholar]
  9. Skurnik M, Wolf-Watz H. Analysis of the yopA gene encoding the Yop1 virulence determinants of Yersinia spp. Mol Microbiol 1989; 3:517–529 [View Article] [PubMed]
     [Google Scholar]
  10. Schütz M, Weiss E-M, Schindler M, Hallström T, Zipfel PF et al. Trimer stability of YadA is critical for virulence of Yersinia enterocolitica. Infect Immun 2010; 78:2677–2690 [View Article] [PubMed]
     [Google Scholar]
  11. Jiang X, Ruiz T, Mintz KP. Characterization of the secretion pathway of the collagen adhesin EmaA of Aggregatibacter actinomycetemcomitans. Mol Oral Microbiol 2012; 27:382–396 [View Article] [PubMed]
     [Google Scholar]
  12. Jiang X, Ruiz T, Mintz KP. The extended signal peptide of the trimeric autotransporter EmaA of Aggregatibacter actinomycetemcomitans modulates secretion. J Bacteriol 2011; 193:6983–6994 [View Article] [PubMed]
     [Google Scholar]
  13. Meuskens I, Saragliadis A, Leo JC, Linke D. Type V secretion systems: an overview of passenger domain functions. Front Microbiol 2019; 10:1163 [View Article] [PubMed]
     [Google Scholar]
  14. Henderson IR, Navarro-Garcia F, Nataro JP. The great escape: structure and function of the autotransporter proteins. Trends Microbiol 1998; 6:370–378 [View Article] [PubMed]
     [Google Scholar]
  15. Linke D, Riess T, Autenrieth IB, Lupas A, Kempf VAJ. Trimeric autotransporter adhesins: variable structure, common function. Trends Microbiol 2006; 14:264–270 [View Article] [PubMed]
     [Google Scholar]
  16. Ackermann N, Tiller M, Anding G, Roggenkamp A, Heesemann J. Contribution of trimeric autotransporter C-terminal domains of oligomeric coiled-coil adhesin (Oca) family members YadA, UspA1, EibA, and Hia to translocation of the YadA passenger domain and virulence of Yersinia enterocolitica. J Bacteriol 2008; 190:5031–5043 [View Article] [PubMed]
     [Google Scholar]
  17. Mühlenkamp M, Oberhettinger P, Leo JC, Linke D, Schütz MS. Yersinia adhesin A (YadA)--beauty & beast. Int J Med Microbiol 2015; 305:252–258 [View Article] [PubMed]
     [Google Scholar]
  18. Tang G, Mintz KP. Glycosylation of the collagen adhesin EmaA of Aggregatibacter actinomycetemcomitans is dependent upon the lipopolysaccharide biosynthetic pathway. J Bacteriol 2010; 192:1395–1404 [View Article] [PubMed]
     [Google Scholar]
  19. Tang G, Ruiz T, Mintz KP. O-polysaccharide glycosylation is required for stability and function of the collagen adhesin EmaA of Aggregatibacter actinomycetemcomitans. Infect Immun 2012; 80:2868–2877 [View Article] [PubMed]
     [Google Scholar]
  20. Watson A, Naughton H, Radermacher M, Mintz KP, Ruiz T. Tomographic analysis of EmaA adhesin glycosylation in Aggregatibacter actinomycetemcomitans. Microsc Microanal 2015; 21:899–900 [View Article]
     [Google Scholar]
  21. Watson A, Tang-Siegel G, Brooks CJ, Radermacher M, Mintz KP et al. Structural significance of EmaA glycosylation in A. actinomycetemcomitans. Microsc Microanal 2016; 22:1132–1133 [View Article]
     [Google Scholar]
  22. Tang G, Ruiz T, Barrantes-Reynolds R, Mintz KP. Molecular heterogeneity of EmaA, an oligomeric autotransporter adhesin of Aggregatibacter (Actinobacillus) actinomycetemcomitans. Microbiology (Reading) 2007; 153:2447–2457 [View Article] [PubMed]
     [Google Scholar]
  23. Perry MB, MacLean LM, Brisson JR, Wilson ME. Structures of the antigenic O-polysaccharides of lipopolysaccharides produced by Actinobacillus actinomycetemcomitans serotypes a, c, d and e. Eur J Biochem 1996; 242:682–688 [View Article] [PubMed]
     [Google Scholar]
  24. Suzuki N, Nakano Y, Yoshida Y, Nezu T, Terada Y et al. Guanosine diphosphate-4-keto-6-deoxy-d-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-d-talose in Actinobacillus actinomycetemcomitans. Eur J Biochem 2002; 269:5963–5971 [View Article] [PubMed]
     [Google Scholar]
  25. Lukomski S, Hoe NP, Abdi I, Rurangirwa J, Kordari P et al. Nonpolar inactivation of the hypervariable streptococcal inhibitor of complement gene (sic) in serotype M1 Streptococcus pyogenes significantly decreases mouse mucosal colonization. Infect Immun 2000; 68:535–542 [View Article] [PubMed]
     [Google Scholar]
  26. Babic A, Guérout A-M, Mazel D. Construction of an improved RP4 (RK2)-based conjugative system. Res Microbiol 2008; 159:545–549 [View Article] [PubMed]
     [Google Scholar]
  27. Mintz KP, Brissette C, Fives-Taylor PM. A recombinase A-deficient strain of Actinobacillus actinomycetemcomitans constructed by insertional mutagenesis using a mobilizable plasmid. FEMS Microbiol Lett 2002; 206:87–92 [View Article] [PubMed]
     [Google Scholar]
  28. Rose JE, Meyer DH, Fives-Taylor PM. Aae, an autotransporter involved in adhesion of Actinobacillus actinomycetemcomitans to epithelial cells. Infect Immun 2003; 71:2384–2393 [View Article] [PubMed]
     [Google Scholar]
  29. Yu C, Ruiz T, Lenox C, Mintz KP. Functional mapping of an oligomeric autotransporter adhesin of Aggregatibacter actinomycetemcomitans. J Bacteriol 2008; 190:3098–3109 [View Article] [PubMed]
     [Google Scholar]
  30. Ruiz T, Mechin I, Bär J, Rypniewski W, Kopperschläger G et al. The 10.8-A structure of Saccharomyces cerevisiae phosphofructokinase determined by cryoelectron microscopy: localization of the putative fructose 6-phosphate binding sites. J Struct Biol 2003; 143:124–134 [View Article] [PubMed]
     [Google Scholar]
  31. Azari F, Radermacher M, Mintz KP, Ruiz T. Correlation of the amino-acid sequence and the 3D structure of the functional domain of EmaA from Aggregatibacter actinomycetemcomitans. J Struct Biol 2012; 177:439–446 [View Article] [PubMed]
     [Google Scholar]
  32. Mintz KP, Fives-Taylor PM. Binding of the periodontal pathogen Actinobacillus actinomycetemcomitans to extracellular matrix proteins. Oral Microbiol Immunol 1999; 14:109–116 [View Article] [PubMed]
     [Google Scholar]
  33. Suzuki N, Nakano Y, Yoshida Y, Nakao H, Yamashita Y et al. Genetic analysis of the gene cluster for the synthesis of serotype a-specific polysaccharide antigen in Aactinobacillus actinomycetemcomitans. Biochim Biophys Acta 2000; 1517:135–138 [View Article] [PubMed]
     [Google Scholar]
  34. Raetz CRH, Whitfield C. Lipopolysaccharide endotoxins. Annu Rev Biochem 2002; 71:635–700 [View Article] [PubMed]
     [Google Scholar]
  35. Whitfield C, Amor PA, Köplin R. Modulation of the surface architecture of gram-negative bacteria by the action of surface polymer:lipid A-core ligase and by determinants of polymer chain length. Mol Microbiol 1997; 23:629–638 [View Article] [PubMed]
     [Google Scholar]
  36. Yu C, Mintz KP, Ruiz T. Investigation of the three-dimensional architecture of the collagen adhesin EmaA of Aggregatibacter actinomycetemcomitans by electron tomography. J Bacteriol 2009; 191:6253–6261 [View Article] [PubMed]
     [Google Scholar]
  37. Roggenkamp A, Ackermann N, Jacobi CA, Truelzsch K, Hoffmann H et al. Molecular analysis of transport and oligomerization of the Yersinia enterocolitica adhesin YadA. J Bacteriol 2003; 185:3735–3744 [View Article] [PubMed]
     [Google Scholar]
  38. Yoshida Y, Nakano Y, Yamashita Y, Koga T. Identification of a genetic locus essential for serotype b-specific antigen synthesis in Actinobacillus actinomycetemcomitans. Infect Immun 1998; 66:107–114 [View Article] [PubMed]
     [Google Scholar]
  39. Kaplan JB, Perry MB, MacLean LL, Furgang D, Wilson ME et al. Structural and genetic analyses of O polysaccharide from Actinobacillus actinomycetemcomitans serotype f. Infect Immun 2001; 69:5375–5384 [View Article] [PubMed]
     [Google Scholar]
  40. Yoshida Y, Nakano Y, Nezu T, Yamashita Y, Koga T. A novel NDP-6-deoxyhexosyl-4-ulose reductase in the pathway for the synthesis of thymidine diphosphate-D-fucose. J Biol Chem 1999; 274:16933–16939 [View Article] [PubMed]
     [Google Scholar]
  41. Perry MB, MacLean LL, Gmür R, Wilson ME. Characterization of the O-polysaccharide structure of lipopolysaccharide from Actinobacillus actinomycetemcomitans serotype b. Infect Immun 1996; 64:1215–1219 [View Article] [PubMed]
     [Google Scholar]
  42. Shibuya N, Amano K, Azuma J, Nishihara T, Kitamura Y et al. 6-Deoxy-D-talan and 6-deoxy-L-talan. Novel serotype-specific polysaccharide antigens from Actinobacillus actinomycetemcomitans. J Biol Chem 1991; 266:16318–16323 [PubMed]
     [Google Scholar]
  43. Zähringer U, Rettenmaier H, Moll H, Senchenkova SN, Knirel YA. Structure of a new 6-deoxy-alpha-D-talan from Burkholderia (Pseudomonas) plantarii strain DSM 6535, which is different from the O-chain of the lipopolysaccharide. Carbohydr Res 1997; 300:143–151 [View Article] [PubMed]
     [Google Scholar]
  44. Gallant CV, Sedic M, Chicoine EA, Ruiz T, Mintz KP. Membrane morphology and leukotoxin secretion are associated with a novel membrane protein of Aggregatibacter actinomycetemcomitans. J Bacteriol 2008; 190:5972–5980 [View Article] [PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.001191
Loading
The serotype a-EmaA adhesin of Aggregatibacter actinomycetemcomitans does not require O-PS synthesis for collagen binding activity
Microbiology 168, 001191 (2022); https://doi.org/10.1099/mic.0.001191
/content/journal/micro/10.1099/mic.0.001191
/content/journal/micro/10.1099/mic.0.001191
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error