Dataset Information

The Hanks-type kinase PknB, targeting the divisome, is not the single player in the Streptococcus thermophilus protein phosphorylation process

ABSTRACT: This work focus on in the role of Ser/Thr/Tyr protein phosphorylation in Streptococcus thermophilus, the only streptococcal species used in food fermentation. During technological processes, S. thermophilus has to adapt to various nutrition and physical-chemical stresses that require rapid adjustments. In this study, we investigated a way for S. thermophilus to regulate specific pathways namely post-translational protein modifications and more specifically protein serine/threonine/tyrosine phosphorylation. We would like to assess the specific role of the only predicted Hanks-type kinase, named PknB. We performed in parallel a global shotgun proteomics and a specific phosphoproteomics analyses on both the wild type strain and its Hanks-type kinase deletion mutant. All the analysis were performed on an Orbitrap Fusion Lumos Tribrid. We showed that the S. thermophilus Ser/Thr/Tyr phosphoproteome is of the same order of magnitude than the other streptococci ones as peptides belonging to 106 proteins (410 phosphopeptides corresponding to 161 peptide sequences with different phosphosite positions) from various metabolic pathways were found phosphorylated in one bacterial growth condition. The phosphorylation occurred for 43 % on serine, 33 % on threonine and 23 % on tyrosine. We demonstrated that the Hanks-type kinase, named PknB in S. thermophilus, targets the divisome and is only one of the players in the Ser/Thr/Tyr phosphorylation process. Consistent with these results, the pknB deletion mutant exhibits a clear phenotype with longer whimsical chains and affected division process.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Streptococcus Thermophilus Lmd-9

SUBMITTER: HENRY CELINE

LAB HEAD: Céline Henry

PROVIDER: PXD011391 | Pride | 2019-07-08

REPOSITORIES: Pride

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Publications

Identification of Hanks-Type Kinase PknB-Specific Targets in the <i>Streptococcus thermophilus</i> Phosphoproteome.

Henry Céline C Haller Lucia L Blein-Nicolas Mélisande M Zivy Michel M Canette Alexis A Verbrugghe Morgane M Mézange Christine C Boulay Mylène M Gardan Rozenn R Samson Samantha S Martin Véronique V André-Leroux Gwenaëlle G Monnet Véronique V

Frontiers in microbiology 20190619

Protein phosphorylation especially on serine/threonine/tyrosine residues are frequent in many bacteria. This post-translational modification has been associated with pathogenicity and virulence in various species. However, only few data have been produced so far on generally recognized as safe bacteria used in food fermentations. A family of kinases known as Hanks-type kinases is suspected to be responsible for, at least, a part of these phosphorylations in eukaryotes as in bacteria. The objecti ...[more]

PMID: 31275266

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Project description:Different transcriptome, metabolome, and phosphoproteome studies have already indicated a regulatory role of Ser/Thr phosphorylation in the central metabolism of S. aureus. However, it remains still unknown how this exactly tunes its metabolism. For that reason, Extreme Pathway Analysis was used to understand metabolic flux regulation by Ser/Thr phosphorylation. YANAsquare software and YANAvergence routine were chosen to calculate pathway activities of WT, ΔpknB, Δstp, and double mutant S. aureus NewmanHG strains. Gene expression data of S. aureus NewmanHG were collected taking the wild type and mutations of kinase (pknB), phosphatase (stp) or both (pknB/stp) phenotypes. The resulting flux changes were modelled with YANAvergence taking the different gene expression data and an established genome scale model of S. aureus metabolic modes as basis. The fluxes of the different strains were next calculated, taking the gene expression data into account. A concerted metabolic change of fluxes was observed. In particular the pathways for peptidoglycan, nucleotide, and aromatic amino acid synthesis as well as catabolism involving aspartate transaminase (GOT) changed significantly compared to the wild type. Single mutations of pknB and stp were compared to illustrate their different contribution to the phenotype, e.g. pyrimidine synthesis is dramatically impaired by pknB deletion but much less by loss of stp. In the double knock out strain, there is higher activity of peptidoglycan, purine, and aromatic amino acids synthesis from glucose, but lower activity of pyrimidine synthesis from glucose compared to WT. The results suggest a probable regulatory role of Ser/Thr phosphorylation in amino acid catabolism and the glycolysis/gluconeogenesis switch to provide the cell with sufficient cell wall components, nucleotides, and aromatic amino acids. Yvck is suggested to serve as regulatory protein linking Ser/Thr phosphorylation to the previous results of S. aureus metabolism. Moreover, Yvck seems in particular to be involved in the switch between glycolysis and gluconeogenesis. However, further studies based on experimental approaches are needed to demonstrate that Ser/Thr phosphorylation and Yvck are necessary for a correct cell growth under gluconeogenic conditions. The metabolic model allows not only a comprehensive representation of these changes for all involved pathways in the four strains, it further points out the function of the previously unannotated ycvk operon.. The relevance of these specific metabolic pathway adaptations regarding the pathogenicity of S.aureus and their contributions to the infection process will be analyzed and discussed.

Dataset Information

The Hanks-type kinase PknB, targeting the divisome, is not the single player in the Streptococcus thermophilus protein phosphorylation process

Publications

Identification of Hanks-Type Kinase PknB-Specific Targets in the <i>Streptococcus thermophilus</i> Phosphoproteome.

Similar Datasets

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