title: Highly conserved nucleotide phosphatase essential for membrane lipid homeostasis in Streptococcus pneumoniae.
abstract: Proteins belonging to the DHH family, a member of the phosphoesterase superfamily, are produced by most bacterial species. While some of these proteins are well studied in Bacillus subtilis and Escherichia coli, their functions in Streptococcus pneumoniae remain unclear. Recently, the highly conserved DHH subfamily 1 protein PapP (SP1298) has been reported to play an important role in virulence. Here, we provide a plausible explanation for the attenuated virulence of the papP mutant. Recombinant PapP specifically hydrolyzed nucleotides 3'-phosphoadenosine-5'-phosphate (pAp) and 5'-phosphoadenylyl-(3'->5')-adenosine (pApA). Deletion of papP, potentially leading to pAp/pApA accumulation, resulted in morphological defects and mis-localization of several cell division proteins. Incubation with both polar solvent and detergent led to robust killing of the papP mutant, indicating that membrane integrity is strongly affected. This is in line with previous studies showing that pAp inhibits the ACP synthase, an essential enzyme involved in lipid precursor production. Remarkably, partial inactivation of the lipid biosynthesis pathway, by inhibition of FabF or depletion of FabH, phenocopied the papP mutant. We conclude that pAp and pApA phosphatase activity of PapP is required for maintenance of membrane lipid homeostasis providing an explanation how inactivation of this protein may attenuate pneumococcal virulence.
authors.affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud Institute for Molecular Life Sciences, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Kuipers
authors.affiliation: Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Synthetic Biology, University of Groningen, Groningen, The Netherlands.
authors.forename: Clement
authors.initials: C
authors.lastname: Gallay
authors.affiliation: Department of Biochemistry, Faculty of Science, Charles University in Prague, Prague, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Martínek
authors.affiliation: Central Facility for Microscopy, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Rohde
authors.affiliation: Department of Biochemistry, Faculty of Science, Charles University in Prague, Prague, Czech Republic.
authors.forename: Markéta
authors.initials: M
authors.lastname: Martínková
authors.affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud Institute for Molecular Life Sciences, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: Samantha L
authors.initials: SL
authors.lastname: van der Beek
authors.affiliation: Department of Molecular Cell Biology, Section Molecular Microbiology, Faculty of Earth and Life Sciences, VU University, Amsterdam, The Netherlands.
authors.forename: Wouter S P
authors.initials: WS
authors.lastname: Jong
authors.affiliation: Center for Molecular and Biomolecular Informatics, Radboud Institute for Molecular Life Sciences, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: Hanka
authors.initials: H
authors.lastname: Venselaar
authors.affiliation: Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Aldert
authors.initials: A
authors.lastname: Zomer
authors.affiliation: Laboratory for Infectious Diseases and Screening, Centre for Infectious Disease Control, RIVM, Bilthoven, The Netherlands.
authors.forename: Hester
authors.initials: H
authors.lastname: Bootsma
authors.affiliation: Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Synthetic Biology, University of Groningen, Groningen, The Netherlands.
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: Veening
authors.affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud Institute for Molecular Life Sciences, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: Marien I
authors.initials: MI
authors.lastname: de Jonge
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D006706:Homeostasis; D008563:Membrane Lipids / Q000378:metabolism*; D009154:Mutation; D009711:Nucleotides / Q000235:genetics / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017384:Sequence Deletion; D013296:Streptococcus pneumoniae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D013329:Structure-Activity Relationship; D014774:Virulence
chemical_list: D000227:Adenine Nucleotides; D001426:Bacterial Proteins; D008563:Membrane Lipids; D009711:Nucleotides; D010744:Phosphoric Monoester Hydrolases; D053487:DEAD-box RNA Helicases