Alginate, inorganic polyphosphate, GTP and ppGpp synthesis co‐regulated in <i>Pseudomonas aeruginosa:</i> implications for stationary phase survival and synthesis of RNA/DNA precursors

Kim, Hong-Yeoul; Schlictman, David; Shankar, Sandeep; Xie, Zhidong; Chakrabarty, Ananda M.; Kornberg, Arthur

doi:10.1046/j.1365-2958.1998.00702.x

Cited by 99 publications

(79 citation statements)

References 30 publications

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“…Deletion of algQ leads to cell death at late exponential phase, a clear consequence of the inability of this mutant to adapt from exponential-phase cell growth to survival mode. This phenotype was rescued by overexpressing AlgQ or Ndk, confirming the role of these proteins in survival adaptation (107).…”

Section: Ppgpp and Pppgppsupporting

confidence: 54%

The Multiple Signaling Systems Regulating Virulence in Pseudomonas aeruginosa

Nadal‐Jimenez

Koch

Thompson

et al. 2012

Microbiol Mol Biol Rev

640

626

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SUMMARY Cell-to-cell communication is a major process that allows bacteria to sense and coordinately react to the fluctuating conditions of the surrounding environment. In several pathogens, this process triggers the production of virulence factors and/or a switch in bacterial lifestyle that is a major determining factor in the outcome and severity of the infection. Understanding how bacteria control these signaling systems is crucial to the development of novel antimicrobial agents capable of reducing virulence while allowing the immune system of the host to clear bacterial infection, an approach likely to reduce the selective pressures for development of resistance. We provide here an up-to-date overview of the molecular basis and physiological implications of cell-to-cell signaling systems in Gram-negative bacteria, focusing on the well-studied bacterium Pseudomonas aeruginosa . All of the known cell-to-cell signaling systems in this bacterium are described, from the most-studied systems, i.e., N -acyl homoserine lactones (AHLs), the 4-quinolones, the global activator of antibiotic and cyanide synthesis (GAC), the cyclic di-GMP (c-di-GMP) and cyclic AMP (cAMP) systems, and the alarmones guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), to less-well-studied signaling molecules, including diketopiperazines, fatty acids (diffusible signal factor [DSF]-like factors), pyoverdine, and pyocyanin. This overview clearly illustrates that bacterial communication is far more complex than initially thought and delivers a clear distinction between signals that are quorum sensing dependent and those relying on alternative factors for their production.

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Section: Ppgpp and Pppgppsupporting

confidence: 54%

The Multiple Signaling Systems Regulating Virulence in Pseudomonas aeruginosa

Nadal‐Jimenez

Koch

Thompson

et al. 2012

Microbiol Mol Biol Rev

640

626

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“…480 aa) with both PMI and GDP-mannose pyrophosphorylase (GMP) (EC 2.7.7.22) activity. An example of the latter type of protein, and one of particular practical importance, is the 56-kDa bifunctional enzyme encoded by algA (179,196,259), which catalyzes the first and third steps in the biosynthesis of alginate, PMI catalyzing the first step (152). This compound is composed of 1,4-linked ␣-Lguluronic acid and ␤-D-mannuronic acid and is of great economic importance, although for commercial production it is usually extracted from marine seaweeds rather than from bacteria (237).…”

Section: Phosphomannose Isomerasesmentioning

confidence: 99%

Microbial Relatives of the Seed Storage Proteins of Higher Plants: Conservation of Structure and Diversification of Function during Evolution of the Cupin Superfamily

Dunwell

Khuri

Gane

2000

Microbiol Mol Biol Rev

313

297

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SUMMARY This review summarizes the recent discovery of the cupin superfamily (from the Latin term “cupa,” a small barrel) of functionally diverse proteins that initially were limited to several higher plant proteins such as seed storage proteins, germin (an oxalate oxidase), germin-like proteins, and auxin-binding protein. Knowledge of the three-dimensional structure of two vicilins, seed proteins with a characteristic β-barrel core, led to the identification of a small number of conserved residues and thence to the discovery of several microbial proteins which share these key amino acids. In particular, there is a highly conserved pattern of two histidine-containing motifs with a varied intermotif spacing. This cupin signature is found as a central component of many microbial proteins including certain types of phosphomannose isomerase, polyketide synthase, epimerase, and dioxygenase. In addition, the signature has been identified within the N-terminal effector domain in a subgroup of bacterial AraC transcription factors. As well as these single-domain cupins, this survey has identified other classes of two-domain bicupins including bacterial gentisate 1,2-dioxygenases and 1-hydroxy-2-naphthoate dioxygenases, fungal oxalate decarboxylases, and legume sucrose-binding proteins. Cupin evolution is discussed from the perspective of the structure-function relationships, using data from the genomes of several prokaryotes, especially Bacillus subtilis. Many of these functions involve aspects of sugar metabolism and cell wall synthesis and are concerned with responses to abiotic stress such as heat, desiccation, or starvation. Particular emphasis is also given to the oxalate-degrading enzymes from microbes, their biological significance, and their value in a range of medical and other applications.

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“…We previously reported that Ndk, which is normally responsible for all NTP formation in the cell, is important for alginate synthesis (24), since it forms complexes with a number of cellular proteins in P. aeruginosa and since such complexes generate predominantly GTP (4,5). It should be noted, however, that GTPgenerating complexes of Ndk with proteins such as pyruvate kinase or Pra (4,5,24) involves the truncated 12-kDa form of Ndk, which is generated uniquely in the mucoid cells by the periplasmic retention of elastase (9,12). An ndk knockout mutant (27), while deficient in alginate synthesis, nevertheless, makes small amounts of alginate.…”

Section: Discussionmentioning

confidence: 99%

“…The Scs protein was shown to copurify with Ndk in P. aeruginosa (10). Ndk exists in two forms in mucoid P. aeruginosa, a cytoplasmic 16-kDa form capable of generating all of the NTPs and a 12-kDa membrane-associated form which forms complexes with pyruvate kinase, Pra, or EF-Tu, and these complexes predominantly use GDP as a substrate, generating GTP (5,12,18). Since Scs copurifies with Ndk, it was of interest to see if Scs modulates Ndk activity in any way.…”

Section: Ndp Kinase Activities Of the Scs With [␥-32 P]atp Or [␥-mentioning

confidence: 99%

Succinyl Coenzyme A Synthetase of Pseudomonas aeruginosa with a Broad Specificity for Nucleoside Triphosphate (NTP) Synthesis Modulates Specificity for NTP Synthesis by the 12-Kilodalton Form of Nucleoside Diphosphate Kinase

2000

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Pseudomonas aeruginosa secretes copious amounts of an exopolysaccharide called alginate during infection in the lungs of cystic fibrosis patients. A mutation in the algR2 gene of mucoid P. aeruginosa is known to exhibit a nonmucoid (nonalginate-producing) phenotype and showed reduced activities of succinyl-coenzyme A (CoA) synthetase (Scs) and nucleoside diphosphate kinase (Ndk), implying coregulation of Ndk and Scs in alginate synthesis. We have cloned and characterized the sucCD operon encoding the ␣ and ␤ subunits of Scs from P. aeruginosa and have studied the role of Scs in generating GTP, an important precursor in alginate synthesis. We demonstrate that, in the presence of GDP, Scs synthesizes GTP using ATP as the phosphodonor and, in the presence of ADP, Scs synthesizes ATP using GTP as a phosphodonor. In the presence of inorganic orthophosphate, succinyl-CoA, and an equimolar amount of ADP and GDP, Scs synthesizes essentially an equimolar amount of ATP and GTP. Such a mechanism of GTP synthesis can be an alternate source for the synthesis of alginate as well as for the synthesis of other macromolecules requiring GTP such as RNA and protein. Scs from P. aeruginosa is also shown to exhibit a broad NDP kinase activity. In the presence of inorganic orthophosphate (P i ), succinyl-CoA, and either GDP, ADP, UDP or CDP, it synthesizes GTP, ATP, UTP, or CTP. Scs was previously shown to copurify with Ndk, presumably as a complex. In mucoid cells of P. aeruginosa, Ndk is also known to exist in two forms, a 16-kDa cytoplasmic form predominant in the log phase and a 12-kDa membrane-associated form predominant in the stationary phase. We have observed that the 16-kDa Ndk-Scs complex present in nonmucoid cells, synthesizes all three of the nucleoside triphosphates from a mixture of GDP, UDP, and CDP, whereas the 12-kDa Ndk-Scs complex specifically present in mucoid cell predominantly synthesizes GTP and UTP but not CTP. Such regulation may promote GTP synthesis in the stationary phase when the bulk of alginate is synthesized by mucoid P. aeruginosa.Our laboratory is interested in studying the regulation of biosynthesis of alginate in Pseudomonas aeruginosa, an opportunistic pathogen. A number of cystic fibrosis (CF) isolates have been studied for their enhanced synthesis and secretion of alginate (16). Alginate is a polymer of D-manuronic and its C-5 epimer L-guluronic acid. GDP-mannuronic acid, derived from GDP-mannose, is the building block of alginate, which requires mannose 1-phosphate and GTP as precursors (17). Thus, alginate synthesis demands a steady supply of GTP for every molecule of mannuronic acid incorporated into a molecule of alginate. Recently, Kamath et al. (9) have shown that the mucoid strains of P. aeruginosa retain an active form of elastase in their periplasm, and this periplasmic elastase acts on 16-kDa Ndk to generate a truncated 12-kDa form that is membrane associated. It has been demonstrated that the membrane-associated 12-kDa nucleoside diphosphate (NDP) kinase (Ndk), in complex with pyruvat...

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Alginate, inorganic polyphosphate, GTP and ppGpp synthesis co‐regulated in Pseudomonas aeruginosa: implications for stationary phase survival and synthesis of RNA/DNA precursors

Cited by 99 publications

References 30 publications

The Multiple Signaling Systems Regulating Virulence in Pseudomonas aeruginosa

The Multiple Signaling Systems Regulating Virulence in Pseudomonas aeruginosa

Microbial Relatives of the Seed Storage Proteins of Higher Plants: Conservation of Structure and Diversification of Function during Evolution of the Cupin Superfamily

Succinyl Coenzyme A Synthetase of Pseudomonas aeruginosa with a Broad Specificity for Nucleoside Triphosphate (NTP) Synthesis Modulates Specificity for NTP Synthesis by the 12-Kilodalton Form of Nucleoside Diphosphate Kinase

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