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Streptococcus pneumoniae detects and responds to foreign bacterial peptide fragments in its environment.

Hathaway LJ, Bättig P, Reber S, Rotzetter JU, Aebi S, Hauser C, Heller M, Kadioglu A, Mühlemann K - Open Biol (2014)

Bottom Line: AliB-like ORF 1 binds specifically peptide SETTFGRDFN, matching 50S ribosomal subunit protein L4 of Enterobacteriaceae, and facilitates upregulation of competence for genetic transformation.We found that AliB-like ORF 2 mediates the early phase of nasopharyngeal colonization in vivo.These findings reveal a completely new concept of pneumococcal interspecies communication which may have implications for communication between other bacterial species and for future interventional therapeutics.

View Article: PubMed Central - PubMed

Affiliation: Institute for Infectious Diseases, University of Bern, Bern, Switzerland.

ABSTRACT
Streptococcus pneumoniae is an important cause of bacterial meningitis and pneumonia but usually colonizes the human nasopharynx harmlessly. As this niche is simultaneously populated by other bacterial species, we looked for a role and pathway of communication between pneumococci and other species. This paper shows that two proteins of non-encapsulated S. pneumoniae, AliB-like ORF 1 and ORF 2, bind specifically to peptides matching other species resulting in changes in the pneumococci. AliB-like ORF 1 binds specifically peptide SETTFGRDFN, matching 50S ribosomal subunit protein L4 of Enterobacteriaceae, and facilitates upregulation of competence for genetic transformation. AliB-like ORF 2 binds specifically peptides containing sequence FPPQS, matching proteins of Prevotella species common in healthy human nasopharyngeal microbiota. We found that AliB-like ORF 2 mediates the early phase of nasopharyngeal colonization in vivo. The ability of S. pneumoniae to bind and respond to peptides of other bacterial species occupying the same host niche may play a key role in adaptation to its environment and in interspecies communication. These findings reveal a completely new concept of pneumococcal interspecies communication which may have implications for communication between other bacterial species and for future interventional therapeutics.

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Identification of AliB-like ORF 1 ligand SETTFGRDFN and AliB-like ORF 2 ligands FPPQSV and FPPQS. (a) AliB-like ORF 1 ligand: extracted ion chromatogram of the doubly charged peptide ion with mass-to-charge ratio (m/z) of 587.26201 (retention time (RT) 47.36 min) with mass tolerance of ±1 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. (b) Representative fragment mass spectrum acquired in the linear trap quadrupole (LTQ) iontrap and sequence interpretation by Phenyx software. Only the y-ion series (fragments appearing to extend from the carboxyl terminus) is annotated. (Individual peaks corresponding to the initial two amino acids S and E at the amino terminus are not visible in the y-ion series and are therefore not annotated but SE is the mass difference between the last detectable y-ion at 957.59 and the singly charged molecular ion of 1173.517.) (c) AliB-like ORF 2 ligands: extracted ion chromatogram of the singly charged peptide ions with mass-to-charge ratio (m/z) of 575.27934 (RT = 45.45 min, corresponding to sequence FPPQS) and 674.34844 (RT = 51.05 min, sequence FPPQSV) with mass tolerance of ±2 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. A representative fragment mass spectrum acquired in Fourier transformation (FT) mode, resolution of 7500, of peptides 575 (d) and 674 (e) is shown. Manual interpretation of the fragment peaks is shown using italic letters for the b-ion (fragments appearing to extend from the amino terminus) and non-italic letters for the y-ion series. Confirmation of correct interpretation was achieved with fragment spectra of synthetic peptides (electronic supplementary material, figure S3).
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RSOB130224F1: Identification of AliB-like ORF 1 ligand SETTFGRDFN and AliB-like ORF 2 ligands FPPQSV and FPPQS. (a) AliB-like ORF 1 ligand: extracted ion chromatogram of the doubly charged peptide ion with mass-to-charge ratio (m/z) of 587.26201 (retention time (RT) 47.36 min) with mass tolerance of ±1 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. (b) Representative fragment mass spectrum acquired in the linear trap quadrupole (LTQ) iontrap and sequence interpretation by Phenyx software. Only the y-ion series (fragments appearing to extend from the carboxyl terminus) is annotated. (Individual peaks corresponding to the initial two amino acids S and E at the amino terminus are not visible in the y-ion series and are therefore not annotated but SE is the mass difference between the last detectable y-ion at 957.59 and the singly charged molecular ion of 1173.517.) (c) AliB-like ORF 2 ligands: extracted ion chromatogram of the singly charged peptide ions with mass-to-charge ratio (m/z) of 575.27934 (RT = 45.45 min, corresponding to sequence FPPQS) and 674.34844 (RT = 51.05 min, sequence FPPQSV) with mass tolerance of ±2 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. A representative fragment mass spectrum acquired in Fourier transformation (FT) mode, resolution of 7500, of peptides 575 (d) and 674 (e) is shown. Manual interpretation of the fragment peaks is shown using italic letters for the b-ion (fragments appearing to extend from the amino terminus) and non-italic letters for the y-ion series. Confirmation of correct interpretation was achieved with fragment spectra of synthetic peptides (electronic supplementary material, figure S3).

Mentions: To find the ligands of S. pneumoniae proteins AliB-like ORF 1 and ORF 2, we expressed recombinant proteins and incubated them in human nasopharyngeal washings and then released bound peptide with urea. LC-MS analysis identified a single peptide ligand for AliB-like ORF 1 with the sequence: SETTFGRDFN (figure 1a,b). For AliB-like ORF 2, two peptide ligands were identified: FPPQSV and FPPQS (figure 1c–e and electronic supplementary material, figure S3). BLAST analysis indicated that AliB-like ORF 1 ligand is in the 50S ribosomal subunit protein L4 of Enterobacteriaceae, including Salmonella enterica, E. coli, Serratia symbiotica and Klebsiella pneumoniae. The sequence of the AliB-like ORF 2 ligand FPPQSV also has multiple bacterial matches including ribosome-associated GTPase EngA of the human commensal Prevotella salivae and sulfate adenylyltransferase of Prevotella tannerae.Figure 1.


Streptococcus pneumoniae detects and responds to foreign bacterial peptide fragments in its environment.

Hathaway LJ, Bättig P, Reber S, Rotzetter JU, Aebi S, Hauser C, Heller M, Kadioglu A, Mühlemann K - Open Biol (2014)

Identification of AliB-like ORF 1 ligand SETTFGRDFN and AliB-like ORF 2 ligands FPPQSV and FPPQS. (a) AliB-like ORF 1 ligand: extracted ion chromatogram of the doubly charged peptide ion with mass-to-charge ratio (m/z) of 587.26201 (retention time (RT) 47.36 min) with mass tolerance of ±1 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. (b) Representative fragment mass spectrum acquired in the linear trap quadrupole (LTQ) iontrap and sequence interpretation by Phenyx software. Only the y-ion series (fragments appearing to extend from the carboxyl terminus) is annotated. (Individual peaks corresponding to the initial two amino acids S and E at the amino terminus are not visible in the y-ion series and are therefore not annotated but SE is the mass difference between the last detectable y-ion at 957.59 and the singly charged molecular ion of 1173.517.) (c) AliB-like ORF 2 ligands: extracted ion chromatogram of the singly charged peptide ions with mass-to-charge ratio (m/z) of 575.27934 (RT = 45.45 min, corresponding to sequence FPPQS) and 674.34844 (RT = 51.05 min, sequence FPPQSV) with mass tolerance of ±2 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. A representative fragment mass spectrum acquired in Fourier transformation (FT) mode, resolution of 7500, of peptides 575 (d) and 674 (e) is shown. Manual interpretation of the fragment peaks is shown using italic letters for the b-ion (fragments appearing to extend from the amino terminus) and non-italic letters for the y-ion series. Confirmation of correct interpretation was achieved with fragment spectra of synthetic peptides (electronic supplementary material, figure S3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043112&req=5

RSOB130224F1: Identification of AliB-like ORF 1 ligand SETTFGRDFN and AliB-like ORF 2 ligands FPPQSV and FPPQS. (a) AliB-like ORF 1 ligand: extracted ion chromatogram of the doubly charged peptide ion with mass-to-charge ratio (m/z) of 587.26201 (retention time (RT) 47.36 min) with mass tolerance of ±1 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. (b) Representative fragment mass spectrum acquired in the linear trap quadrupole (LTQ) iontrap and sequence interpretation by Phenyx software. Only the y-ion series (fragments appearing to extend from the carboxyl terminus) is annotated. (Individual peaks corresponding to the initial two amino acids S and E at the amino terminus are not visible in the y-ion series and are therefore not annotated but SE is the mass difference between the last detectable y-ion at 957.59 and the singly charged molecular ion of 1173.517.) (c) AliB-like ORF 2 ligands: extracted ion chromatogram of the singly charged peptide ions with mass-to-charge ratio (m/z) of 575.27934 (RT = 45.45 min, corresponding to sequence FPPQS) and 674.34844 (RT = 51.05 min, sequence FPPQSV) with mass tolerance of ±2 ppm from the negative control (i) and positive sample (ii), intensity scale set at the same level. A representative fragment mass spectrum acquired in Fourier transformation (FT) mode, resolution of 7500, of peptides 575 (d) and 674 (e) is shown. Manual interpretation of the fragment peaks is shown using italic letters for the b-ion (fragments appearing to extend from the amino terminus) and non-italic letters for the y-ion series. Confirmation of correct interpretation was achieved with fragment spectra of synthetic peptides (electronic supplementary material, figure S3).
Mentions: To find the ligands of S. pneumoniae proteins AliB-like ORF 1 and ORF 2, we expressed recombinant proteins and incubated them in human nasopharyngeal washings and then released bound peptide with urea. LC-MS analysis identified a single peptide ligand for AliB-like ORF 1 with the sequence: SETTFGRDFN (figure 1a,b). For AliB-like ORF 2, two peptide ligands were identified: FPPQSV and FPPQS (figure 1c–e and electronic supplementary material, figure S3). BLAST analysis indicated that AliB-like ORF 1 ligand is in the 50S ribosomal subunit protein L4 of Enterobacteriaceae, including Salmonella enterica, E. coli, Serratia symbiotica and Klebsiella pneumoniae. The sequence of the AliB-like ORF 2 ligand FPPQSV also has multiple bacterial matches including ribosome-associated GTPase EngA of the human commensal Prevotella salivae and sulfate adenylyltransferase of Prevotella tannerae.Figure 1.

Bottom Line: AliB-like ORF 1 binds specifically peptide SETTFGRDFN, matching 50S ribosomal subunit protein L4 of Enterobacteriaceae, and facilitates upregulation of competence for genetic transformation.We found that AliB-like ORF 2 mediates the early phase of nasopharyngeal colonization in vivo.These findings reveal a completely new concept of pneumococcal interspecies communication which may have implications for communication between other bacterial species and for future interventional therapeutics.

View Article: PubMed Central - PubMed

Affiliation: Institute for Infectious Diseases, University of Bern, Bern, Switzerland.

ABSTRACT
Streptococcus pneumoniae is an important cause of bacterial meningitis and pneumonia but usually colonizes the human nasopharynx harmlessly. As this niche is simultaneously populated by other bacterial species, we looked for a role and pathway of communication between pneumococci and other species. This paper shows that two proteins of non-encapsulated S. pneumoniae, AliB-like ORF 1 and ORF 2, bind specifically to peptides matching other species resulting in changes in the pneumococci. AliB-like ORF 1 binds specifically peptide SETTFGRDFN, matching 50S ribosomal subunit protein L4 of Enterobacteriaceae, and facilitates upregulation of competence for genetic transformation. AliB-like ORF 2 binds specifically peptides containing sequence FPPQS, matching proteins of Prevotella species common in healthy human nasopharyngeal microbiota. We found that AliB-like ORF 2 mediates the early phase of nasopharyngeal colonization in vivo. The ability of S. pneumoniae to bind and respond to peptides of other bacterial species occupying the same host niche may play a key role in adaptation to its environment and in interspecies communication. These findings reveal a completely new concept of pneumococcal interspecies communication which may have implications for communication between other bacterial species and for future interventional therapeutics.

Show MeSH
Related in: MedlinePlus