title: Characterization of pneumococcal Ser/Thr protein phosphatase phpP mutant and identification of a novel PhpP substrate, putative RNA binding protein Jag.
abstract: BACKGROUND
  Reversible protein phosphorylation catalyzed by protein kinases and phosphatases is the primary mechanism for signal transduction in all living organisms. Streptococcus pneumoniae encodes a single Ser/Thr protein kinase, StkP, which plays a role in virulence, stress resistance and the regulation of cell wall synthesis and cell division. However, the role of its cognate phosphatase, PhpP, is not well defined.
  
  
  RESULTS
  Here, we report the successful construction of a ΔphpP mutant in the unencapsulated S. pneumoniae Rx1 strain and the characterization of its phenotype. We demonstrate that PhpP negatively controls the level of protein phosphorylation in S. pneumoniae both by direct dephosphorylation of target proteins and by dephosphorylation of its cognate kinase, StkP. Catalytic inactivation or absence of PhpP resulted in the hyperphosphorylation of StkP substrates and specific phenotypic changes, including sensitivity to environmental stresses and competence deficiency. The morphology of the ΔphpP cells resembled the StkP overexpression phenotype and conversely, overexpression of PhpP resulted in cell elongation mimicking the stkP null phenotype. Proteomic analysis of the phpP knock-out strain permitted identification of a novel StkP/PhpP substrate, Spr1851, a putative RNA-binding protein homologous to Jag. Here, we show that pneumococcal Jag is phosphorylated on Thr89. Inactivation of jag confers a phenotype similar to the phpP mutant strain.
  
  
  CONCLUSIONS
  Our results suggest that PhpP and StkP cooperatively regulate cell division of S. pneumoniae and phosphorylate putative RNA binding protein Jag.
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Aleš
authors: A
authors: Ulrych
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Nela
authors: N
authors: Holečková
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Jana
authors: J
authors: Goldová
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic. linda@biomed.cas.cz.
authors: Linda
authors: http://orcid.org/0000-0003-3433-0262
authors: L
authors: Doubravová
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Oldřich
authors: O
authors: Benada
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Olga
authors: O
authors: Kofroňová
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic.
authors: Petr
authors: P
authors: Halada
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic. branny@biomed.cas.cz.
authors: Pavel
authors: P
authors: Branny
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D002473:Cell Wall / Q000378:metabolism; D055786:Gene Knockout Techniques; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology; D010641:Phenotype; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D015398:Signal Transduction; D013296:Streptococcus pneumoniae / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D050505:Mutant Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D017346:Protein Serine-Threonine Kinases; D010749:Phosphoprotein Phosphatases
keywords: Cell division; Jag; Phosphorylation; Protein kinase; Protein phosphatase; Signal transduction; Streptococcus