Growth of <i>Pseudomonas aeruginosa</i> in zinc poor environments is promoted by a nicotianamine‐related metallophore

Mastropasqua, Maria Chiara; D’Orazio, Melania; Cerasi, Mauro; Pacello, Francesca; Gismondi, Angelo; Canini, Antonella; Canuti, Lorena; Consalvo, Ada; Ciavardelli, Domenico; Chirullo, Barbara; Pasquali, Paolo; Battistoni, Andrea

doi:10.1111/mmi.13834

Cited by 89 publications

(153 citation statements)

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“…Staphylopine could transport copper, nickel, cobalt, zinc and iron [5,21,22], and participates in zinc uptake in zinc-scarce environments [21,23]. Pseudopaline is involved in nickel uptake in minimal media, whereas it is responsible for zinc uptake in zinc-scarce environment [6,24]. For both S. aureus and P. aeruginosa , literature reports a link between the production of opine-type metallophores and bacterial infection.…”

Section: Introductionmentioning

confidence: 99%

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Laffont

Brutesco

Hajjar

et al. 2019

Preprint

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3In metal-scarce environments, some pathogenic bacteria produce opine-type metallophores 1 4 mainly to face the host's nutritional immunity. This is the case of staphylopine, pseudopaline and 1 5 yersinopine, identified in Staphylococcus aureus, Pseudomonas aeruginosa and Yersinia pestis 1 6respectively. These metallophores are synthesized by two (CntLM) or three enzymes (CntKLM), 1 7 CntM catalyzing the last step of biosynthesis using diverse substrates (pyruvate or α -ketoglutarate), 1 8 pathway intermediates (xNA or yNA) and cofactors (NADH or NADPH), depending on the species. 1 9Here, we explored substrate specificity of CntM by combining bioinformatics and structural analysis 2 0 with chemical synthesis and enzymatic studies. We found that NAD(P)H selectivity was mainly due to 2 1 the amino acid at position 33 (S. aureus numbering) which ensures a preferential binding to NADPH 2 2 when it is an arginine. Moreover, whereas CntM from P. aeruginosa preferentially uses yNA over 2 3 xNA, the staphylococcal enzyme is not stereospecific. Most importantly, selectivity towards α -2 4 ketoacids is largely governed by a single residue at position 150 of CntM (S. aureus numbering): an 2 5 aspartate at this position ensures selectivity towards pyruvate whereas an alanine leads to the 2 6 consumption of both pyruvate and α -ketoglutarate. Modifying this residue in P. aeruginosa led to a 2 7complete reversal of selectivity. Thus, opine-type metallophore diversity is mainly mediated by the 2 8absence/presence of a cntK gene encoding a histidine racemase, and the presence of an 2 9 aspartate/alanine at position 150 of CntM. These two simple rules predict the production of a fourth 3 0 metallophore by Paenibacillus mucilaginosus, which was confirmed in vitro and called bacillopaline. 3 1 3 2 3 3 3 5metals required for their growth. This is particularly the case for pathogenic bacteria that have to 3 6 confront the host's immune system. Indeed, the so-called "nutritional immunity" induces an additional 3 7 metal limitation by sequestering iron, zinc or manganese to prevent bacterial growth [1-4]. To face 3 8 this metal restriction, bacteria have developed metallophores to recover metals. In this context, 3 9 nicotianamine-like metallophores have been identified in some bacteria as playing an important role in 4 0 metal acquisition strategies. Staphylopine, pseudopaline and yersinopine are the three examples 4 1 2 currently known and recently identified in S. aureus [5], P. aeruginosa [6,7] and Y. pestis [8] 4 2respectively. The biosynthesis of these nicotianamine-like metallophores occurs in two or three steps 4 3 depending on the species. When it is present, as in S. aureus, CntK, a histidine racemase, transforms 4 4

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Section: Introductionmentioning

confidence: 99%

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Laffont

Brutesco

Hajjar

et al. 2019

Preprint

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“…To maintain zinc homeostasis, bacterial species have evolved multiple systems to import and export zinc in environments of zinc limitation or excess, respectively (33, 34). In P. aeruginosa , the two best-studied zinc acquisition systems are the high-affinity ZnuABC system (35–37) and the pseudopaline system (38, 39). ZnuABC is a member of the ATP-binding cassette (ABC) transporters, consisting of a zinc-specific binding periplasmic protein (ZnuA), an inner membrane permease (ZnuB), and an ATPase (ZnuC) (35, 37, 40).…”

Section: Introductionmentioning

confidence: 99%

“…Loss of ZnuA in P. aeruginosa PAO1 results in an ~60% reduction in cellular zinc accumulation (35). The pseudopaline system is primarily involved in zinc uptake in zinc poor environments and zinc transport into the cell is achieved via the action of a four-gene operon ( cntOLMI , also termed zrmABCD ) (38, 39). This operon includes the genes for a TonB-dependent outer membrane pseudopaline receptor (CntO), two biosynthetic enzymes (CntL and CntM) responsible for synthesizing pseudopaline and an inner membrane transporter (CntI) involved in pseudopaline secretion (36, 38, 39).…”

Section: Introductionmentioning

confidence: 99%

“…The pseudopaline system is primarily involved in zinc uptake in zinc poor environments and zinc transport into the cell is achieved via the action of a four-gene operon ( cntOLMI , also termed zrmABCD ) (38, 39). This operon includes the genes for a TonB-dependent outer membrane pseudopaline receptor (CntO), two biosynthetic enzymes (CntL and CntM) responsible for synthesizing pseudopaline and an inner membrane transporter (CntI) involved in pseudopaline secretion (36, 38, 39). Both the ZnuABC system and the pseudopaline operon are negatively regulated by zinc level through the Fur-like zinc uptake regulator Zur (35, 37, 39), which can sense and respond to femtomolar changes of cytosolic zinc concentrations (41).…”

Section: Introductionmentioning

confidence: 99%

“…This operon includes the genes for a TonB-dependent outer membrane pseudopaline receptor (CntO), two biosynthetic enzymes (CntL and CntM) responsible for synthesizing pseudopaline and an inner membrane transporter (CntI) involved in pseudopaline secretion (36, 38, 39). Both the ZnuABC system and the pseudopaline operon are negatively regulated by zinc level through the Fur-like zinc uptake regulator Zur (35, 37, 39), which can sense and respond to femtomolar changes of cytosolic zinc concentrations (41). When zinc is bound, Zur represses transcription of the zinc uptake systems and loss of Zur results in higher cytoplasmic zinc in P. aeruginosa (36, 37).…”

Section: Introductionmentioning

confidence: 99%

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Availability of Zinc Impacts Interactions BetweenStreptococcus sanguinisandPseudomonas aeruginosain Co-culture

Gifford

Hampton

et al. 2019

Preprint

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23Airway infections associated with cystic fibrosis (CF) are polymicrobial. We reported 24 previously that clinical isolates of P. aeruginosa promote the growth of a variety of 25 streptococcal species. To explore the mechanistic basis of this interaction, we performed a 26 genetic screen to identify mutants of Streptococcus sanginuis SK36 whose growth was no 27 longer enhanced by P. aeruginosa PAO1. Mutations in zinc uptake systems of S. sanginuis 28 SK36 reduced growth of these strains by 1-3 log compared to wild-type S. sanginuis SK36 29 when grown in coculture with P. aeruginosa PA01, while exogenous zinc (0.1-10 μm) rescued 30 the coculture defect of zinc uptake mutants of S. sanginuis SK36. Zinc uptake mutants of S. 31 sanginuis SK36 had no obvious growth defect in monoculture. Consistent with a competition 32 for zinc driving coculture dynamics, S. sanginuis SK36 grown in coculture with P. aeruginosa 33 showed increased expression of zinc uptake genes compared to S. sanginuis grown alone. 34 Strains of P. aeruginosa PAO1 defective in zinc transport also supported more robust growth 35 by S. sanginuis compared to coculture with wild-type P. aeruginosa PAO1. An analysis of 118 36 CF sputum samples revealed that total zinc levels varied from ~5-145 μM. At relatively low 37 zinc levels, Pseudomonas and Streptococcus were found in approximately equal abundance; 38 at higher zinc levels, we observed an increasing relative abundance of Pseudomonas and 39 decline of Streptococcus, perhaps as a result of increasing zinc toxicity. Together, our data 40 indicate that the relative abundance of these microbes in the CF airway may be impacted by 41 zinc levels. 42 IMPORTANCE. Polymicrobial infections in CF likely impact patient health, but the 43 mechanism(s) underlying such interactions are poorly understood. Here we show that 44 interactions between Pseudomonas and Streptococcus are modulated by zinc availability 45 using an in vitro model system, and clinical data are consistent with this model. Together with 46 previous studies, our work supports a role for metal homeostasis as a key factor driving 47 49Cystic fibrosis (CF) is a monogenic autosomal recessive disorder caused by mutations in the 50 cystic fibrosis transmembrane conductance regulator (CFTR) gene (1). It is estimated that 51 ~70,000 individuals in the world are affected by CF and the most common mutation, caused 52 by a deletion of phenylalanine at the 508th amino acid within the CFTR protein (ΔF508), is 53 found in approximately 70% of this population (2, 3). CFTR dysfunction affects several body 54 systems, and progressive lung disease due to chronic and recurrent microbial infections is 55 the leading cause of morbidity and mortality in individuals with CF (4, 5). It has been shown 56 that CF airway infections are polymicrobial (6, 7), and the composition and interspecies 57 interactions within the polymicrobial communities can have profound and diverse 58 consequences, including on bacterial growth (8-10), as well as disease progression and 59 thera...

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Function‐Oriented Natural Product Synthesis

Chen

Lei

2021

Chin. J. Chem.

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What is the most favorite and original chemistry developed in your research group? By far the most original and groundbreaking work we have done is the discovery of the first naturally occurring TRUE Diels-Alderase which shows tremendous synthetic potential in biocatalysis for efficiently making structurally complex and bioactive molecules. How do you get into this specific field? Could you please share some experiences with our readers? I have a great passion in synthetic chemistry, and I am always fascinated by making organic molecules that may help us explore new biology and develop medicine for human diseases. Therefore, my research programs are relatively broad, spanning from natural product synthesis, chemical biology, synthetic biology, biocatalysis to drug discovery. What is the most important personality for scientific research? In my personal opinion, curiosity, ambition, and dedication are the most important personalities for scientific research. What is your favorite journal(s)? My favorite journals are: 1) Cell, when you read a Cell paper, you can get a full story of the original discovery and understand the detailed logic on how the research is planned and conducted; 2) Accounts of Chemical Research, this journal provides a short review on how a chemist establishes a specific new research field, very inspiring! What's your hobbies? I love many sports, soccer, American football, basketball, etc. I am also good at Seal Cutting, one of the four major traditional Chinese arts. If you have anything else to tell our readers, please feel free to do so. I would like to share one advice with the young scientists who just start their independent research career: BE DIFFERENT from your Ph.D. and Postdoc advisors, try to establish your own research direction! It's always easy to quickly get paper published when you take advantage of your advisor's research system, but it's very challenging to establish your own reputation and prove your originality in this way.

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Growth of Pseudomonas aeruginosa in zinc poor environments is promoted by a nicotianamine‐related metallophore

Cited by 89 publications

References 70 publications

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Availability of Zinc Impacts Interactions BetweenStreptococcus sanguinisandPseudomonas aeruginosain Co-culture

Function‐Oriented Natural Product Synthesis

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