1887

Abstract

is a major human pathogen responsible for significant mortality and morbidity worldwide. Within the annotated genome of the pneumococcus lies a previously uncharacterized protein tyrosine phosphatase which shows homology to low molecular weight protein tyrosine phosphatases (LMWPTPs). LMWPTPs modulate many processes critical for the pathogenicity of a number of bacteria including capsular polysaccharide biosynthesis, stress response and persistence in host macrophages. Here, we demonstrate that Spd1837 is indeed a LMWPTP, by purifying the protein, and characterizing its phosphatase activity. Spd1837 showed specific tyrosine phosphatase activity, and it did not form higher order oligomers in contrast to many other LMWPTPs. Substrate-trapping assays using the wild-type and the phosphatase-deficient Spd1837 identified potential substrates/interacting proteins including major metabolic enzymes such as ATP-dependent-6-phosphofructokinase and Hpr kinase/phosphorylase. Given the tight association between the bacterial basic physiology and virulence, this study hopes to prompt further investigation of how the pneumococcus controls its metabolic flux via the LMWPTP Spd1837.

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2018-04-01
2024-03-28
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References

  1. Morona JK, Morona R, Paton JC. Attachment of capsular polysaccharide to the cell wall of Streptococcus pneumoniae type 2 is required for invasive disease. Proc Natl Acad Sci USA 2006; 103:8505–8510 [View Article][PubMed]
    [Google Scholar]
  2. Morona JK, Morona R, Miller DC, Paton JC. Streptococcus pneumoniae capsule biosynthesis protein CpsB is a novel manganese-dependent phosphotyrosine-protein phosphatase. J Bacteriol 2002; 184:577–583 [View Article][PubMed]
    [Google Scholar]
  3. Bender MH, Yother J. CpsB is a modulator of capsule-associated tyrosine kinase activity in Streptococcus pneumoniae . J Biol Chem 2001; 276:47966–47974 [View Article][PubMed]
    [Google Scholar]
  4. Standish AJ, Whittall JJ, Morona R. Tyrosine phosphorylation enhances activity of pneumococcal autolysin LytA. Microbiology 2014; 160:2745–2754 [View Article][PubMed]
    [Google Scholar]
  5. Nourikyan J, Kjos M, Mercy C, Cluzel C, Morlot C et al. Autophosphorylation of the bacterial tyrosine-kinase CpsD connects capsule synthesis with the cell cycle in Streptococcus pneumoniae . PLoS Genet 2015; 11:e1005518 [View Article][PubMed]
    [Google Scholar]
  6. Standish AJ, Morona R. The role of bacterial protein tyrosine phosphatases in the regulation of the biosynthesis of secreted polysaccharides. Antioxid Redox Signal 2014; 20:2274–2289 [View Article][PubMed]
    [Google Scholar]
  7. Ahmad Z, Harvey RM, Paton JC, Standish AJ, Morona R. Role of Streptococcus pneumoniae OM001 operon in capsular polysaccharide production, virulence and survival in human saliva. PLoS One 2018; 13:e0190402 [View Article][PubMed]
    [Google Scholar]
  8. Klein G, Dartigalongue C, Raina S. Phosphorylation-mediated regulation of heat shock response in Escherichia coli . Mol Microbiol 2003; 48:269–285 [View Article][PubMed]
    [Google Scholar]
  9. Musumeci L, Bongiorni C, Tautz L, Edwards RA, Osterman A et al. Low-molecular-weight protein tyrosine phosphatases of Bacillus subtilis . J Bacteriol 2005; 187:4945–4956 [View Article][PubMed]
    [Google Scholar]
  10. Mukhopadhyay A, Kennelly PJ. A low molecular weight protein tyrosine phosphatase from Synechocystis sp. strain PCC 6803: enzymatic characterization and identification of its potential substrates. J Biochem 2011; 149:551–562 [View Article][PubMed]
    [Google Scholar]
  11. Romero P, López R, García E. Key role of amino acid residues in the dimerization and catalytic activation of the autolysin LytA, an important virulence factor in Streptococcus pneumoniae . J Biol Chem 2007; 282:17729–17737 [View Article][PubMed]
    [Google Scholar]
  12. Standish AJ, Salim AA, Capon RJ, Morona R. Dual inhibition of DNA polymerase PolC and protein tyrosine phosphatase CpsB uncovers a novel antibiotic target. Biochem Biophys Res Commun 2013; 430:167–172 [View Article][PubMed]
    [Google Scholar]
  13. Blanchetot C, Chagnon M, Dubé N, Hallé M, Tremblay ML. Substrate-trapping techniques in the identification of cellular PTP targets. Methods 2005; 35:44–53 [View Article][PubMed]
    [Google Scholar]
  14. Cathro P, McCarthy P, Hoffmann P, Zilm P. Isolation and identification of Enterococcus faecalis membrane proteins using membrane shaving, 1D SDS/PAGE, and mass spectrometry. FEBS Open Bio 2016; 6:586–593 [View Article][PubMed]
    [Google Scholar]
  15. Morona JK, Paton JC, Miller DC, Morona R. Tyrosine phosphorylation of CpsD negatively regulates capsular polysaccharide biosynthesis in Streptococcus pneumoniae . Mol Microbiol 2000; 35:1431–1442 [View Article][PubMed]
    [Google Scholar]
  16. Salomone-Stagni M, Musiani F, Benini S. Characterization and 1.57 Å resolution structure of the key fire blight phosphatase AmsI from Erwinia amylovora . Acta Crystallogr F Struct Biol Commun 2016; 72:903–910 [View Article][PubMed]
    [Google Scholar]
  17. Tabernero L, Evans BN, Tishmack PA, van Etten RL, Stauffacher CV. The structure of the bovine protein tyrosine phosphatase dimer reveals a potential self-regulation mechanism. Biochemistry 1999; 38:11651–11658[PubMed] [Crossref]
    [Google Scholar]
  18. Akerud T, Thulin E, van Etten RL, Akke M. Intramolecular dynamics of low molecular weight protein tyrosine phosphatase in monomer-dimer equilibrium studied by NMR: a model for changes in dynamics upon target binding. J Mol Biol 2002; 322:137–152 [View Article][PubMed]
    [Google Scholar]
  19. Bernadó P, Akerud T, García de La Torre J, Akke M, Pons M. Combined use of NMR relaxation measurements and hydrodynamic calculations to study protein association. Evidence for tetramers of low molecular weight protein tyrosine phosphatase in solution. J Am Chem Soc 2003; 125:916–923 [View Article][PubMed]
    [Google Scholar]
  20. Nath S, Banerjee R, Sen U. Atomic resolution crystal structure of VcLMWPTP-1 from Vibrio cholerae O395: insights into a novel mode of dimerization in the low molecular weight protein tyrosine phosphatase family. Biochem Biophys Res Commun 2014; 450:390–395 [View Article][PubMed]
    [Google Scholar]
  21. Andrade A, Valvano MA. A Burkholderia cenocepacia gene encoding a non-functional tyrosine phosphatase is required for the delayed maturation of the bacteria-containing vacuoles in macrophages. Microbiology 2014; 160:1332–1345 [View Article][PubMed]
    [Google Scholar]
  22. Wong D, Bach H, Sun J, Hmama Z, Av-Gay Y. Mycobacterium tuberculosis protein tyrosine phosphatase (PtpA) excludes host vacuolar-H+-ATPase to inhibit phagosome acidification. Proc Natl Acad Sci USA 2011; 108:19371–19376 [View Article][PubMed]
    [Google Scholar]
  23. Geno KA, Hauser JR, Gupta K, Yother J. Streptococcus pneumoniae phosphotyrosine phosphatase CpsB and alterations in capsule production resulting from changes in oxygen availability. J Bacteriol 2014; 196:1992–2003 [View Article][PubMed]
    [Google Scholar]
  24. Kobir A, Shi L, Boskovic A, Grangeasse C, Franjevic D et al. Protein phosphorylation in bacterial signal transduction. Biochim Biophys Acta 2011; 1810:989–994 [View Article][PubMed]
    [Google Scholar]
  25. Soufi B, Gnad F, Jensen PR, Petranovic D, Mann M et al. The Ser/Thr/Tyr phosphoproteome of Lactococcus lactis IL1403 reveals multiply phosphorylated proteins. Proteomics 2008; 8:3486–3493 [View Article][PubMed]
    [Google Scholar]
  26. Sun X, Ge F, Xiao CL, Yin XF, Ge R et al. Phosphoproteomic analysis reveals the multiple roles of phosphorylation in pathogenic bacterium Streptococcus pneumoniae . J Proteome Res 2010; 9:275–282 [View Article][PubMed]
    [Google Scholar]
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