New structural and functional defects in polyphosphate deficient bacteria: a cellular and proteomic study.
BACKGROUND
Inorganic polyphosphate (polyP), a polymer of tens or hundreds of phosphate residues linked by ATP-like bonds, is found in all organisms and performs a wide variety of functions. PolyP is synthesized in bacterial cells by the actions of polyphosphate kinases (PPK1 and PPK2) and degraded by exopolyphosphatase (PPX). Bacterial cells with polyP deficiencies due to knocking out the ppk1 gene are affected in many structural and important cellular functions such as motility, quorum sensing, biofilm formation and virulence among others. The cause of this pleiotropy is not entirely understood.


RESULTS
The overexpression of exopolyphosphatase in bacteria mimicked some pleitropic defects found in ppk1 mutants. By using this approach we found new structural and functional defects in the polyP-accumulating bacteria Pseudomonas sp. B4, which are most likely due to differences in the polyP-removal strategy. Colony morphology phenotype, lipopolysaccharide (LPS) structure changes and cellular division malfunction were observed. Finally, we used comparative proteomics in order to elucidate the cellular adjustments that occurred during polyP deficiency in this bacterium and found some clues that helped to understand the structural and functional defects observed.


CONCLUSIONS
The results obtained suggest that during polyP deficiency energy metabolism and particularly nucleoside triphosphate (NTP) formation were affected and that bacterial cells overcame this problem by increasing the flux of energy-generating metabolic pathways such as tricarboxilic acid (TCA) cycle, beta-oxidation and oxidative phosphorylation and by reducing energy-consuming ones such as active transporters and amino acid biosynthesis. Furthermore, our results suggest that a general stress response also took place in the cell during polyP deficiency.
Varela
Cristian
C
Department of Biology, Faculty of Sciences, Laboratory of Molecular Microbiology and Biotechnology & Millennium Institute of Cell Dynamics and Biotechnology, University of Chile, Las Palmeras 3425, Nuñoa, Santiago, Chile.
Mauriaca
Cecilia
C
Paradela
Alberto
A
Albar
Juan P
JP
Jerez
Carlos A
CA
Chávez
Francisco P
FP
D017766:Acid Anhydride Hydrolases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms; D004734:Energy Metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D011122:Polyphosphates / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*
D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D011122:Polyphosphates; D020543:Proteome; D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase; D017766:Acid Anhydride Hydrolases; C020137:exopolyphosphatase