Importance of protein Ser/Thr/Tyr phosphorylation for bacterial pathogenesis

Køhler, Julie Bonne; Jers, Carsten; Senissar, Mériem; Shi, Lei; Derouiche, Abderahmane; Mijaković, Ivan

doi:10.1002/1873-3468.13797

Cited by 32 publications

(35 citation statements)

References 264 publications

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“…This will facilitate functional interpretations and guide experimental design. In this regard, future automated integration of phospho data sets with structural databases or prediction tools will be of great advantage (42).…”

Section: Journal Of Lipid Research Volume 61 2020mentioning

confidence: 99%

The protein kinase PknB negatively regulates biosynthesis and trafficking of mycolic acids in mycobacteria

Le¹,

Locard‐Paulet²,

Stella³

et al. 2020

Journal of Lipid Research

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Mycobacterium tuberculosis is the causative agent of tuberculosis and remains one of the most widespread and deadliest bacterial pathogens in the world. A distinguishing feature of mycobacteria that sets them apart from other bacteria is the unique architecture of their cell wall, characterized by various species-specific lipids, most notably mycolic acids (MAs). Therefore, targeted inhibition of enzymes involved in MA biosynthesis, transport, and assembly has been extensively explored in drug discovery. Additionally, more recent evidence suggests that many enzymes in the MA biosynthesis pathway are regulated by kinase-mediated phosphorylation, thus opening additional drug development opportunities. However, how phosphorylation regulates MA production remains unclear. Here, we employed genetic strategies combined with lipidomics and phosphoproteomics approaches to investigate the role of protein phosphorylation in Mycobacterium. The results of this analysis revealed that the Ser/Thr protein kinase PknB regulates export of MAs and promotes remodeling of the mycobacterial cell envelope. In particular, we identified the essential mycobacterial membrane protein large 3 (MmpL3) as a substrate negatively regulated by PknB. Taken together, our findings add to the understanding of how PknB activity affects the mycobacterial MA biosynthesis pathway and reveal the essential role of protein phosphorylation/dephosphorylation in governing lipid metabolism, paving the way towards novel antimycobacterial strategies.

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Section: Journal Of Lipid Research Volume 61 2020mentioning

confidence: 99%

The protein kinase PknB negatively regulates biosynthesis and trafficking of mycolic acids in mycobacteria

Le¹,

Locard‐Paulet²,

Stella³

et al. 2020

Journal of Lipid Research

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show abstract

“…Part of the reason for the limited overlap in previous experiments may have been the diversity in enrichment methods and experimental protocols, limited sensitivity of the workflows and variability in sampling method and timepoint. Our workflow allows the reliable identification of a large set of phosphoproteins in comparison with other studies (11,12,16,17). Moreover, we applied rigorous quality control of phosphosite assignment.…”

Section: Discussionmentioning

confidence: 99%

“…In bacteria, phosphorylation has been implicated in diverse processes such as cell cycle regulation, cellular differentiation, morphogenesis, and metabolism, persistence and virulence (1). Whereas phosphoproteomics initially focused on model organisms such as the Gram-negative Escherichia coli (14) and Gram-positive Bacillus subtilis (15), phosphoproteomes have been determined for a large number of pathogens as well (16,17).…”

Section: Introductionmentioning

confidence: 99%

“…Together, these findings suggest that targeting phosphorylation might be a viable strategy for novel antimicrobial or anti-virulence strategies (16,18). Despite the wealth of phosphoproteomic studies (16,17), phosphorylation in Clostridia is largely unexplored. A single study has been performed in Clostridium acetobutylicum, a solventogenic species with biotechnological relevance, that identified a total of 61 phosphorylated proteins (19).…”

Section: Introductionmentioning

confidence: 99%

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Clostridioides difficile phosphoproteomics shows an expansion of phosphorylated proteins in stationary growth phase

Smits

Mohammed

et al. 2021

Preprint

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Phosphorylation is a post-translational modification that can affect both house-keeping functions and virulence characteristics in bacterial pathogens. In the Gram-positive enteropathogen Clostridioides difficile the extent and nature of phosphorylation events is poorly characterized, though a protein-kinase mutant strain demonstrates pleiotropic phenotypes. Here, we used an immobilized metal affinity chromatography strategy to characterize serine, threonine and tyrosine phosphorylation in C. difficile. We find limited protein phosphorylation in the exponential growth phase but a sharp increase in the number of phosphopeptides after the onset of stationary growth phase. Among the overall more than 1500 phosphosites, our approach identifies expected targets and phosphorylation sites, including the protein kinase PrkC, the anti-sigma-F factor antagonist (SpoIIAA), the anti-sigma-B factor antagonist (RsbV) and HPr kinase/phosphorylase (HprK). Analysis of high-confidence phosphosites shows that phosphorylation on serine residues is most common, followed by threonine and tyrosine phosphorylation. This work forms the basis for a further investigation into the contributions of individual kinases to the overall phosphoproteome of C. difficile and the role of phosphorylation in C. difficile physiology and pathogenesis.ImportanceIn this manuscript, we present a comprehensive analysis of protein phosphorylation in the Gram-positive enteropathogen Clostridioides difficile. To date, only limited evidence on the role of phosphorylation in regulation in this organism has been published; the current study is expected to form the basis for research on this post-translational modification in C. difficile.

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“…These kinases and other types of protein kinases form a complex phosphorylation network in prokaryotes ( Jers et al, 2008 ). Although Ser/Thr protein kinases are not DNA-binding proteins, they can mediate the phosphorylation of substrate proteins to regulate gene expression, affect the cellular localization, metabolism and many other cellular functions ( Bonne Køhler et al, 2020 ). For example, the Ser/Thr protein kinase PrkC in Bacillus anthracis can mediate the expression of the metabolic protein Eno, which acts as an intrinsic memory controller and affects the germination process, thereby helping B. anthracis survive the nutritional shift and promoting the role of glycolytic enzymes in the carbohydrate metabolic process during germination ( Virmani et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Impacts of Ser/Thr Protein Kinase Stk1 on the Proteome, Twitching Motility, and Competitive Advantage in Pseudomonas aeruginosa

et al. 2021

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Pseudomonas aeruginosa is a ubiquitous gram-negative bacterium in the environment and a leading cause of nosocomial infections worldwide. Therefore, it is listed by the WHO as a human pathogen that urgently needs the development of new antibacterial drugs. Recent findings have demonstrated that eukaryote-type Ser/Thr protein kinases play a vital role in regulating various bacterial physiological processes by catalyzing protein phosphorylation. Stk1 has proven to be a Ser/Thr protein kinase in P. aeruginosa. However, the regulatory roles of Stk1 have not yet been revealed. Thus, we constructed a stk1 knockout mutant (∆stk1) from the P. aeruginosa PAO1 strain and employed a Tandem Mass Tag (TMT) labeling-based quantitative proteomic strategy to characterize proteome-wide changes in response to the stk1 knockout. In total, 620 differentially expressed proteins, among which 288 proteins were upregulated and 332 proteins were downregulated, were identified in ∆stk1 compared with P. aeruginosa PAO1. A detailed bioinformatics analysis of these differentially expressed proteins was performed, including GO annotation, protein domain profile, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, subcellular localization and enrichment analysis. Notably, the downregulation of type IV pilus-related proteins and upregulation of T6SS-H1-related proteins were found in the ∆stk1 strain, and the results were corroborated by quantitative PCR at the mRNA level. Further experiments confirmed that the loss of stk1 weakens bacterial twitching motility and promotes a growth competition advantage, which are, respectively, mediated by type IV pilus-related proteins and T6SS-H1-related proteins. These findings contribute to a better understanding of the physiological role of Stk1, and proteomic data will help further investigations of the roles and mechanisms of Stk1 in P. aeruginosa, although the detailed regulation and mechanism of Stk1 still need to be revealed.

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Importance of protein Ser/Thr/Tyr phosphorylation for bacterial pathogenesis

Cited by 32 publications

References 264 publications

The protein kinase PknB negatively regulates biosynthesis and trafficking of mycolic acids in mycobacteria

The protein kinase PknB negatively regulates biosynthesis and trafficking of mycolic acids in mycobacteria

Clostridioides difficile phosphoproteomics shows an expansion of phosphorylated proteins in stationary growth phase

Impacts of Ser/Thr Protein Kinase Stk1 on the Proteome, Twitching Motility, and Competitive Advantage in Pseudomonas aeruginosa

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