Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

E. coli toxin YjjJ (HipH) is a Ser/Thr protein kinase that impacts cell division, carbon metabolism and ribosome assembly

ABSTRACT: Protein Ser/Thr kinases are post-translational regulators of key molecular processes in bacteria, such as cell division and antibiotic tolerance. Here we characterize the E. coli toxin YjjJ (HipH), a putative protein kinase annotated as a member of the family of HipA-like Ser/Thr kinases which are involved in antibiotic tolerance. Using SILAC-based phosphoproteomics we provide experimental evidence that YjjJ is a Ser/Thr protein kinase and its primary protein substrates are the ribosomal protein RpmE (L31) and the carbon storage regulator CsrA. YjjJ activity impacts ribosome assembly, cell division and central carbon metabolism but it does not increase antibiotic tolerance as does its homologue HipA. Intriguingly, overproduction of YjjJ and its kinase-deficient variant can activate HipA and other kinases, pointing to a crosstalk between Ser/Thr kinases in E. coli.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Escherichia Coli

SUBMITTER: Nicolas Nalpas

LAB HEAD: Boris Macek

PROVIDER: PXD033071 | Pride | 2022-11-28

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

High persister protein kinase HipA and its gain-of-function variant HipA7 phosphorylate different substrate pools in Escherichia coli

Project description:Bacterial persistence, the ability to survive antibiotic treatment by entering a physiologically dormant state, is a serious biomedical problem. Protein Ser/Thr kinase HipA, the first toxin connected to bacterial multidrug tolerance (persistence), exerts its function by phosphorylating glutamate--tRNA ligase GltX, leading to a halt in translation, accumulation of ppGpp and induction of persistence. Intriguingly, its variant HipA7 is able to induce significantly higher levels of persistence despite being less toxic for the cell. We postulated that this phenotypic difference may be driven by diverse substrate pools of the two kinase variants. To address this, we ectopically expressed hipA and hipA7 in E. coli and monitored their in vivo substrates during growth inhibition and resuscitation using SILAC-based quantitative phosphoproteomics. Our assays confirmed that both forms of the kinase phosphorylate GltX as the main substrate. Importantly, HipA phosphorylated several additional substrates involved in protein synthesis, transcription and replication, such as ribosomal protein L11 and SeqA. Conversely, HipA7 had a lower kinase activity, no additional substrates under tested conditions and showed a similar substrate pool only when expressed at significantly higher levels. The two forms of the kinase also differed in autophosphorylation level, which was significantly lower in HipA7. Initial testing of the novel HipA substrate L11 did not reveal a connection between HipA-induced phosphorylation and RelA-dependent persistence, opening the possibility that the novel HipA substrates are predominantly responsible for the toxic phenotype of the kinase. Our results contribute to understanding of HipA7 action and present a resource for studies of HipA-related persistence.

2018-09-17 | PXD009173 | Pride

RNA_Seq-based global transcriptome analysis of S. pneumoniae mutants lacking PhpP and StkP

Project description:We report the mRNA expression profile of Streptococcus pneumoniae Type 2 D39 strain-derived mutant lacking the phpP gene encoding eukaryote-type ser/thr phosphatase and the stkP gene encoding Ser/Thr protein kinase.

2019-04-13 | GSE113337 | GEO

Substrate selectivity for semisynthetic CK2 proteins with Pin1

Project description:Protein Ser/Thr kinase CK2 is involved in a myriad of cellular processes including cell growth and proliferation by phosphorylating hundreds of substrates, yet the regulation process of CK2 function is poorly understood. The CK2 catalytic subunit, CK2α, is phosphorylated at Thr344 and phosphorylation on the C-terminal tail of CK2α is required for interaction with Pin1 protein. The substrate selectivity for protein kinase CK2α was examined by performing kinase assays on protein microarrays spotted with 17,000 human proteins. Semisynthetic CK2α proteins were prepared to contain an unmodified C-terminal tail or phospho-Thr (pThr) at T344. These semisynthetic proteins were used to determine if the phosphorylation-dependent interaction of CK2α with Pin1 can modulate the substrate selectivity for CK2. The different semisynthetic CK2α proteins (unmodified and pThr344) were tested alone and in the presence of the recombinant Pin1 protein. Pin1 has been shown to interaction with CK2α only when CK2α is phoshorylated on its C-terminal site (including Thr344). In the study presented here, kinase assays were performed using two different semisynthetic CK2α proteins: unmodified C-terminal tail and phospho-Thr (pThr) at 344. The semisynthetic proteins were each tested alone and in the presence of the recombinant Pin1 protein. There were four different kinase conditions and each condition was performed in duplicate.

2011-10-24 | E-GEOD-33150 | biostudies-arrayexpress

Proteomic profiling of human monocytes stimulated in vitro with N. meningitidis

Project description:Meningococcal sepsis is an overwhelming form of the sepsis syndrome which may cause mortality within 12-24 hours in previously healthy children and adults, where the causative infectious agent is N. meningitidis, an obligate human pathogen. The genomic changes induced by N. meningitidis are modulated by the anti-inflammatory cytokine interleukin-10 (IL-10), which is present in large quantities in plasma from patients with meningococcal sepsis. This present study investigated kinase activities in human monocytes stimulated by N. meningitidis and IL-10. The first aim was to identify array peptides that could indicate which signaling pathways were activated or inhibited by the host response to the meningococci. The second aim was to detect whether IL-10 affected N. meningitidis-nduced phosphorylation of array peptides, in order to identify potential targets of the IL-10 anti-inflammatory response. We approached this using a strategy where elutriation-purified human monocytes are stimulated in vitro with N. meningitidis and IL-10, with concentrations corresponding to previously measured levels in patients with fulminant meningococcal septicemia. This work examined activation or inhibition of signaling pathways mediated by tyrosine kinases when purified human monocytes are in vitro incubated with N. meningitidis in the presence or absence of IL-10.

2017-12-15 | E-MTAB-4795 | biostudies-arrayexpress

PrkA regulates Bacillus subtilis sporulation through a Lon protease activity

Project description:In Bacillus subtilis, sporulation is a sequential and highly regulated process. Phosphorylation events by Histidine or Serine/Threonine kinases are key points in this regulation. PrkA has been proposed to be an essential Serine kinase for the initiation of sporulation but its kinase activity has not been clearly demonstrated so far. Indeed, neither its autophosphorylation nor identification of a B. subtilis phosphorylated substrate was unambiguously established. Bioinformatic homology searches revealed sequence similarities with the AAA+ ATP-dependent Lon protease family. Here, we showed that PrkA is indeed able to hydrolyse the α-casein, an exogenous substrate of Lon proteases, in an ATP-dependent manner. We also showed that this ATP-dependent protease activity is essential for PrkA function in sporulation since mutation in the Walker A motif leads to a sporulation defect. Furthermore, we found that PrkA protease activity is tightly regulated by phosphorylation events involving one of the Ser/Thr kinases of B. subtilis, PrkC, as characterized by mass spectrometry from 3 in vitro independent experiments (1: PrkA+PrkC/2:PrKA-S219E+PrkC/3: PrKA-T217E+PrkC). We finally demonstrated that PrkA regulation of the transcriptional factor σK via the transition phase regulator ScoC is certainly indirect.

2022-10-15 | PXD032931 | Pride

Calcium/calmodulin dependent protein kinase kinase 2 (CAMKK2), an alternative to curcumin in head and neck squamous cell carcinoma.

Project description:Curcumin has been shown to exhibit anti-neoplastic effects. However, due to its poor bioavailability, use of curcumin as an anti-cancer drug is limited. It is thus necessary to identify molecules that could be targeted using anti-cancer agents as alternatives to curcumin. For this, it is important to understand the underlying signaling pathways mediating the anti-neoplastic effects of curcumin. We carried out phosphoproteomic analysis (TiO2-based enrichment of Ser/Thr kinases) of head and neck cancer cell line, CAL27 treated with curcumin to identify curcumin mediated signaling pathways. This resulted in the identification of 5921 phosphopeptides (phosphosite probability≥75%) corresponding 1,878 proteins. Of these, 275 and 335 phosphopeptides corresponding to 183 and 242 proteins (≥2.0-fold) were found to be hyper- and hypo-phosphorylated respectively, in response to curcumin treatment

2022-03-03 | PXD007256 | Pride

RNA-seq analysis of S. pneumoniae Pn-STP (PhpP) mutants

Project description:We report the mRNA expression profile of Streptococcus pneumoniae Type 2 D39 strain-derived mutant lacking the phpP gene encoding eukaryote-type ser/thr phosphatase .

2016-10-27 | GSE74421 | GEO

Transcription profiling of atm mutant, adm mutant and wild type whole plants and roots of Arabdiposis after gamma ray irradiation in a time series

Project description:Whole seedlings of wild type (4d) and atm mutants (4d) have been analyzed after a gamma ray irradiation of 0.75h, 1.5h, 3h & 5h (time course). Roots of wt (4d), atm (3d) and atr (4d) mutants have been analyzed after a 1h irradiation.<br><br> Ataxia Telangiectasia Mutated (ATM), encodes a large protein with a phosphatidylinositol 3-kinase (PI3K)-like domain at the C terminus (reviewed by Rotman and Shiloh, 1998). PI3K-related proteins make up a large family of Ser-Thr protein kinases, numerous members of which are involved in the regulation of cell cycle progression, responses to DNA damage, and the maintenance of genomic stability (Hoekstra, 1997). AtATM plays an essential role in meiosis and in the somatic response to DNA damage in plants, similar to the function of ATM in mammals and other eukaryotes.<br>Ataxia telangiectasia-mutated and Rad3-related (ATR) plays a central role in cell-cycle regulation, transmitting DNA damage signals to downstream effectors of cell-cycle progression.

2006-10-30 | E-MEXP-780 | biostudies-arrayexpress

Master mitotic kinases regulate viral genome delivery during papillomavirus cell entry

Project description:Cell cycle progression into mitosis induce cellular rearrangements such as mitotic spindle formation, Golgi fragmentation, and nuclear envelope breakdown. Like certain retroviruses, nuclear delivery of HPV16 genomes is facilitated by these processes during entry into host cells by tethering of the viral DNA to mitotic chromosomes through the minor capsid protein L2. However, the mechanism of delivery onto and tethering to the condensed chromosomes is barely understood on a mechanistic level. To date it is unclear, which cellular proteins facilitate this process in interaction with L2 or how this process is regulated. Here, we discovered that HPV16 minor capsid protein L2 is phosphorylated during entry upon mitosis onset on conserved residues within the chromosome-binding region (CBR) that is responsible for nuclear import. The crucial L2 phosphorylations occurred sequentially by the master mitotic kinases CDK1 and PLK1. L2 phosphorylation, thus, not only regulated timely delivery of HPV16 vDNA to mitotic chromatin at mitosis onset, but also likely resulted in a conformational switch in L2 that allowed engagement of cellular proteins for this purpose. In summary, our work demonstrates for the first time a crucial role of mitotic kinases for nuclear entry of a DNA virus and provides important insights into the molecular mechanism of pathogen import into the nucleus during mitosis.

2022-12-22 | PXD028704 | Pride

Proteomic investigation uncovers potential targets and target sites of pneumococcal serine-threonine kinase StkP and phosphatase PhpP

Project description:Like eukaryotes, different bacterial species express one or more Ser/Thr kinases and phosphatases that operate in various signaling networks by catalyzing phosphorylation and dephosphorylation of proteins that can immediately regulate biochemical pathways by altering protein function. The human pathogen Streptococcus pneumoniae encodes a single Ser/Thr kinase-phosphatase couple known as StkP-PhpP, which has shown to be crucial in the regulation of cell wall synthesis and cell division. In this study, we applied proteomics to further understand the physiological role of pneumococcal PhpP and StkP with an emphasis on phosphorylation events on Ser and Thr residues. Therefore, the proteome of the non-encapsulated D39 strain (WT), a kinase (ΔstkP) and phosphatase mutant (ΔphpP) were compared in a mass spectrometry based label-free quantification experiment. Results show that a loss of function of PhpP causes an increased abundance of proteins in the phosphate uptake system Pst. Quantitative proteomic data demonstrated an effect of StkP and PhpP on the two-component systems ComDE, LiaRS, CiaRH and VicRK. To obtain further information on the function, targets and target sites of PhpP and StkP we combined the advantages of phosphopeptide enrichment using titan dioxide and spectral library based data evaluation for sensitive detection of changes in the phosphoproteome of the wild type and the mutant strains. According to the role of StkP in cell division we identified several proteins involved in cell wall synthesis and cell division that are apparently phosphorylated by StkP. Unlike StkP, the physiological function of the co-expressed PhpP is poorly understood. For the first time we were able to provide a list of previously unknown putative targets of PhpP. Under these new putative targets of PhpP are, among others, five proteins with direct involvement in cell division (DivIVA, GpsB) and peptidoglycan biosynthesis (MltG, MreC, MacP).

2020-01-22 | PXD015268 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data