A Novel Mode of Regulation of the Staphylococcus aureus Catabolite Control Protein A (CcpA) Mediated by Stk1 Protein Phosphorylation

Leiba, Jade; Hartmann, Torsten; Cluzel, Marie-Eve; Cohen‐Gonsaud, Martin; Delolme, Frédéric; Bischoff, Markus; Molle, Virginie

doi:10.1074/jbc.m112.418913

Cited by 64 publications

(77 citation statements)

References 49 publications

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“…To identify regulatory proteins essential for NOṙ esistance in S. aureus, we screened a library of transposon mutants with insertions in every nonessential predicted transcriptional regulator for signs of NO˙sensitivity (mutants obtained from the Nebraska Transposon Mutant Library). In total, we screened 115 regulatory transposon mutants comprising 110 transcriptional regulators (approximately 80% of all predicted transcriptional regulators in S. aureus), three sigma factors, and a Ser/ Thr kinase/phosphatase enzyme pair known to modulate the functions of multiple regulatory proteins (44)(45)(46)(47)(48). To screen for NO˙sensitivity, we compared the normalized lag time (see Materials and Methods) of the regulatory mutants to that of WT S. aureus following treatment with NO˙at the time of inoculation.…”

Section: Resultsmentioning

confidence: 99%

Regulatory Requirements for Staphylococcus aureus Nitric Oxide Resistance

et al. 2016

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The ability of Staphylococcus aureus to resist host innate immunity augments the severity and pervasiveness of its pathogenesis. Nitric oxide (NO˙) is an innate immune radical that is critical for the efficient clearance of a wide range of microbial pathogens. Exposure of microbes to NO˙typically results in growth inhibition and induction of stress regulons. S. aureus, however, induces a metabolic state in response to NO˙that allows for continued replication and precludes stress regulon induction. The regulatory factors mediating this distinctive response remain largely undefined. Here, we employ a targeted transposon screen and transcriptomics to identify and characterize five regulons essential for NO˙resistance in S. aureus: three virulence regulons not formerly associated with NO˙resistance, SarA, CodY, and Rot, as well as two regulons with established roles, Fur and SrrAB. We provide new insights into the contributions of Fur and SrrAB during NO˙stress and show that the S. aureus ⌬sarA mutant, the most sensitive of the newly identified mutants, exhibits metabolic dysfunction and widespread transcriptional dysregulation following NO˙exposure. Altogether, our results broadly characterize the regulatory requirements for NO˙resistance in S. aureus and suggest an intriguing overlap between the regulation of NO˙resistance and virulence in this well-adapted human pathogen. IMPORTANCEThe prolific human pathogen Staphylococcus aureus is uniquely capable of resisting the antimicrobial radical nitric oxide (NO˙), a crucial component of the innate immune response. However, a complete understanding of how S. aureus regulates an effective response to NO˙is lacking. Here, we implicate three central virulence regulators, SarA, CodY, and Rot, as major players in the S. aureus NO˙response. Additionally, we elaborate on the contribution of two regulators, SrrAB and Fur, already known to play a crucial role in S. aureus NO˙resistance. Our study sheds light on a unique facet of S. aureus pathogenicity and demonstrates that the transcriptional response of S. aureus to NO˙is highly pleiotropic and intrinsically tied to metabolism and virulence regulation. The versatile Gram-positive pathogen Staphylococcus aureus is the leading cause of skin and soft tissue infections (SSTIs) in the United States but can also cause more severe illnesses, including pneumonia, osteomyelitis, endocarditis, and bacteremia (1-5). The ability of S. aureus to infect virtually every tissue of the body can be partially attributed to its extensive capacity for subverting host immunity. One unique defense of S. aureus against the immune system is resistance to nitric oxide (NO˙), a membrane-permeable radical and broad-spectrum innate immune effector required for the clearance of bacterial, viral, and fungal pathogens (6). The ability of S. aureus to continue growth in the presence of NO˙at concentrations that are inhibitory to other bacteria, including closely related staphylococci, contributes to its invasiveness and pathogenic success. For example...

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Section: Resultsmentioning

confidence: 99%

Regulatory Requirements for Staphylococcus aureus Nitric Oxide Resistance

et al. 2016

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“…Leiba et al went on to show affected phenotypes through Thr18/33 regulatory control, including formation of biofilms in vitro. 97 Interestingly, despite the exceptional conservation of Thr18/33 across multiple bacterial species, CcpA orthologs are unable to accept a phosphate by their native eSTKs, however cross-species phosphorylation is possible. 97 Considering the high homology of eSTKs and CcpA orthologs across Gram-positive species, these surprising results suggest that slight changes within amino acid composition might afford a rapid reactivation of 'dormant' eSTK controlled regulatory circuits.…”

Section: Staphylococcus Aureusmentioning

confidence: 99%

“…97 Interestingly, despite the exceptional conservation of Thr18/33 across multiple bacterial species, CcpA orthologs are unable to accept a phosphate by their native eSTKs, however cross-species phosphorylation is possible. 97 Considering the high homology of eSTKs and CcpA orthologs across Gram-positive species, these surprising results suggest that slight changes within amino acid composition might afford a rapid reactivation of 'dormant' eSTK controlled regulatory circuits. How such subtle discrimination by eSTKs between homologous variants of CcpA and other eSTKregulated substrates is possible when all possess the conserved phosphorylatable residues remains an exciting topic of future research.…”

Section: Staphylococcus Aureusmentioning

confidence: 99%

“…These residues are located within the CcpA DNA-binding domain and are predicted to hydrogen bond with target DNA. 97 In this example, eSTK phosphorylation of CcpA affords the bacteria a simple additional mechanism of regulatory control, whereby protein-DNA interactions can be directly abrogated by the PTMs to allow activation of CcpA-repressed catabolite promoters. Leiba et al went on to show affected phenotypes through Thr18/33 regulatory control, including formation of biofilms in vitro.…”

Section: Staphylococcus Aureusmentioning

confidence: 99%

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Regulation of transcription by eukaryotic-like serine-threonine kinases and phosphatases in Gram-positive bacterial pathogens

Wright

Ulijasz

2014

Virulence

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“…These residues are situated in the CcpA DNA-binding domain and are presumed to interact with the target DNA by forming hydrogen bonds. CcpA phosphorylation by Stk1 abrogates the protein-DNA interaction, and leads to activation of CcpA-repressed promoters implicated in sugar metabolism and biofilm formation (Leiba et al, 2012) (Fig. 1a).…”

Section: Serine/threonine Phosphorylation Of Bacterial Trsmentioning

confidence: 99%

Serine/threonine/tyrosine phosphorylation regulates DNA binding of bacterial transcriptional regulators

et al. 2015

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Reversible phosphorylation of bacterial transcriptional regulators (TRs) belonging to the family of two-component systems (TCSs) is a well-established mechanism for regulating gene expression. Recent evidence points to the fact that reversible phosphorylation of bacterial TRs on other types of residue, i.e. serine, threonine, tyrosine and cysteine, is also quite common. The phosphorylation of the ester type (phospho-serine/threonine/tyrosine) is more stable than the aspartate phosphorylation of TCSs. The kinases which catalyse these phosphorylation events (Hanks-type serine/threonine protein kinases and bacterial protein tyrosine kinases) are also much more promiscuous than the TCS kinases, i.e. each of them can phosphorylate several substrate proteins. As a consequence, the dynamics and topology of the signal transduction networks depending on these kinases differ significantly from the TCSs. Here, we present an overview of different classes of bacterial TR phosphorylated and regulated by serine/threonine and tyrosine kinases. Particular attention is given to examples when serine/threonine and tyrosine kinases interact with TCSs, phosphorylating either the histidine kinases or the response regulators. We argue that these promiscuous kinases connect several signal transduction pathways and serve the role of signal integration. Bacterial two-component systems (TCSs)TCSs are signal transduction devices that were initially discovered in bacteria (Ninfa & Magasani, 1986;Nixon et al., 1986). They play an important role in signal sensing and response to various stimuli, enabling the organisms to adapt to environmental changes. A typical TCS consists of a histidine kinase (HK) and a corresponding response regulator (RR) (Stock et al., 2000;Gao & Stock, 2009). HK usually possesses a highly variable sensor domain and a conserved kinase core. Following environmental stimulus, a signal ligand binds to the sensor domain and results in the autophosphorylation of the kinase core at a conserved histidine residue, at the expense of ATP. Next, the phosphoryl group is transferred from HK to a conserved aspartate in the regulatory domain of the RR. RRs usually contain two domains: a regulatory domain with the conserved phosphorylatable aspartate and a variable effector domain. Phosphorylation activates the effector domain of RRs, triggering the physiological response. As phosphohistidine has a very short half-life in aqueous solutions at neutral pH, in the absence of the environmental signal the system switches itself off very rapidly.Many effector domains of bacterial RRs have DNA-binding capacity. This allows RRs to function as transcriptional regulators (TRs) and consequently change gene transcription when they become phosphorylated. In Escherichia coli, osmoregulation of porin proteins OmpF and OmpC is under transcriptional control of the TCS EnvZ/OmpR. The phosphorylation of OmpR by EnvZ changes its affinity for the promoter region of ompF and ompC, resulting in different transcriptional levels of these genes (Forst et al., 1...

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A Novel Mode of Regulation of the Staphylococcus aureus Catabolite Control Protein A (CcpA) Mediated by Stk1 Protein Phosphorylation

Cited by 64 publications

References 49 publications

Regulatory Requirements for Staphylococcus aureus Nitric Oxide Resistance

Regulatory Requirements for Staphylococcus aureus Nitric Oxide Resistance

Regulation of transcription by eukaryotic-like serine-threonine kinases and phosphatases in Gram-positive bacterial pathogens

Serine/threonine/tyrosine phosphorylation regulates DNA binding of bacterial transcriptional regulators

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