Dataset Information

Targets of kinase/phosphatase signalling pair StkP/PhpP in S. pneumoniae and their involvement in response to cell wall stress

ABSTRACT: Streptococcus pneumoniae is an opportunistic human pathogen which encodes a single eukaryotic-type Ser/Thr protein kinase StkP and its functional counterpart protein phosphatase PhpP. These signalling enzymes play a crucial role in the coordination of cell division and growth in pneumococcus. In this study, we determined the proteomic and phosphoproteomic profiles of relevant mutants. A comparison with the wild type provided a representative data set of new phosphoacceptor sites and StkP-dependent substrates. StkP phosphorylates key proteins involved in cell division and cell wall biosynthesis in unencapsulated laboratory Rx1 strain as well as encapsulated virulent strain D39. Moreover, we show that StkP plays important role in eliciting an adaptive response induced by cell wall directed antibiotic. Phosphorylation of WalK sensor kinase and decline of the abundance of WalK and proteins of core WalR/K regulon implies cross-talk between StkP and WalR/K two-component system. Inspection of proteome profiles revealed gene clusters regulated by catabolite control mechanism suggesting a tight coupling of carbon flow and cell wall homeostasis. The disbalance of steady-state protein phosphorylation in the mutants as well as after antibiotic treatment is accompanied by accumulation of global Spx regulator indicating Spx-mediated envelope stress response. In summary, StkP translates the sensed signal about the cell-wall status to key cell division and regulatory proteins and in this way controls cell cycle and cell wall homeostasis.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Streptococcus Pneumoniae Serotype 2 (strain D39 / Nctc 7466) Streptococcus Pneumoniae (strain Atcc Baa-255 / R6) Streptococcus Pneumoniae

SUBMITTER: Ivo Fabrik

LAB HEAD: Pavel Branny

PROVIDER: PXD024033 | Pride | 2022-05-12

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20190103_IF022_Sp10_A_01.raw	Raw
	20190103_IF022_Sp10_A_02.raw	Raw
	20190103_IF022_Sp10_B_01.raw	Raw
	20190103_IF022_Sp10_B_02.raw	Raw
	20190103_IF022_Sp10_C_01.raw	Raw

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Publications

Cell Wall Stress Stimulates the Activity of the Protein Kinase StkP of Streptococcus pneumoniae, Leading to Multiple Phosphorylation.

Ulrych Aleš A Fabrik Ivo I Kupčík Rudolf R Vajrychová Marie M Doubravová Linda L Branny Pavel P

Journal of molecular biology 20211021 24

Streptococcus pneumoniae is an opportunistic human pathogen that encodes a single eukaryotic-type Ser/Thr protein kinase StkP and its functional counterpart, the protein phosphatase PhpP. These signaling enzymes play critical roles in coordinating cell division and growth in pneumococci. In this study, we determined the proteome and phosphoproteome profiles of relevant mutants. Comparison of those with the wild-type provided a representative dataset of novel phosphoacceptor sites and StkP-depend ...[more]

PMID: 34688688

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Project description:Abstract of associated manuscript: The Bacillus subtilis extracytoplasmic function (ECF) sigma(M) factor is activated by cell envelope stress elicited by antibiotics, and by acid, heat, ethanol and superoxide stresses. Here, we have used several complementary approaches to identify genes controlled by sigma(M). In many cases, expression is only partially dependent on sigma(M) because of both overlapping promoter recognition with other ECF sigma factors and the presence of additional promoter elements. Genes regulated by sigma(M) have a characteristic pattern of induction in response to cell envelope-acting antibiotics as evidenced by hierarchical clustering analysis. sigma(M) also contributes to the expression of the Spx transcription factor and thereby indirectly regulates genes of the Spx regulon. Cell envelope stress responses also include regulons controlled by sigma(W), sigma(B) and several two-component regulatory systems (e.g. LiaRS, YycFG, BceRS). Activation of the sigma(M) regulon increases expression of proteins functioning in transcriptional control, cell wall synthesis and shape determination, cell division, DNA damage monitoring, recombinational repair and detoxification. WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van). Each experiment was conducted at least twice using two independent total RNA preparations. For vancomycin untreated and treated experiments, untreated samples were labeled with Alexa Fluor 555 and treated samples with Alexa Fluor 647. For WT vs. mutant experiments, wild type was labeled with Alexa Fluor 555 and mutants with Alexa Fluor 647. For dye swap experiment, wild-type was labeled with Alexa Fluor 647 and mutant with Alexa Fluor 555. Bacillus subtilis CU1065, WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van)