Evolutionary aspects of urea utilization by fungi

Navarathna, Dhammika H. M. L. P.; Harris, Steven D.; Roberts, D. A.; Nickerson, Kenneth W.

doi:10.1111/j.1567-1364.2009.00602.x

Cited by 43 publications

(52 citation statements)

References 26 publications

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“…In this study, we demonstrate that coexpression of the S. pombe high affinity Ni 2 þ -transporter gene (Nic1; Eitinger et al, 2000) was required for Ure2-dependent growth of S. cerevisiae at low Ni 2 þ concentrations. This is in line with phylogenetic research, which indicates that acquisition of the bi-functional ATP-dependent Dur1,2 and loss of ATP-independent urease in the ancestor of Saccharomycotina yeasts coincided with the loss of the high affinity nickel transporter (Navarathna et al, 2010;Zhang et al, 2009).…”

Section: Expression Of a Ni-dependent Atp-independent Urease In S Cesupporting

confidence: 80%

“…S. cerevisiae) (Kurtzman and Robnett, 2013;Weiss et al, 2013) harbour the DUR1,2 gene encoding ATP-dependent urease. It has been hypothesized that the evolutionary advantage of expending ATP was to allow yeasts such as S. cerevisiae to eliminate all nickelrequiring reactions, thus reducing the number of transition metals for which cellular homoeostasis and regulation is required (Navarathna et al, 2010). In this study, we demonstrate that coexpression of the S. pombe high affinity Ni 2 þ -transporter gene (Nic1; Eitinger et al, 2000) was required for Ure2-dependent growth of S. cerevisiae at low Ni 2 þ concentrations.…”

Section: Expression Of a Ni-dependent Atp-independent Urease In S Cementioning

confidence: 90%

“…To this end we sought to replace the native ATP-dependent urea assimilation gene DUR1,2 with the complete ATP-independent urea assimilation system from S. pombe (Bacanamwo et al, 2002;Eitinger et al, 2000;Mobley et al, 1995;Navarathna et al, 2010). The functionality of the heterologous urease was characterized in vivo and in vitro.…”

Section: Introductionmentioning

confidence: 99%

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Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

Milne

Luttik

Cueto-Rojas

et al. 2015

Metabolic Engineering

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a b s t r a c tIn microbial processes for production of proteins, biomass and nitrogen-containing commodity chemicals, ATP requirements for nitrogen assimilation affect product yields on the energy producing substrate. In Saccharomyces cerevisiae, a current host for heterologous protein production and potential platform for production of nitrogen-containing chemicals, uptake and assimilation of ammonium requires 1 ATP per incorporated NH 3 . Urea assimilation by this yeast is more energy efficient but still requires 0.5 ATP per NH 3 produced. To decrease ATP costs for nitrogen assimilation, the S. cerevisiae gene encoding ATP-dependent urease (DUR1,2) was replaced by a Schizosaccharomyces pombe gene encoding ATP-independent urease (ure2), along with its accessory genes ureD, ureF and ureG. Since S. pombe ure2 is a Ni 2 þ -dependent enzyme and Saccharomyces cerevisiae does not express native Ni 2 þ -dependent enzymes, the S. pombe high-affinity nickel-transporter gene (nic1) was also expressed. Expression of the S. pombe genes into dur1,2Δ S. cerevisiae yielded an in vitro ATPindependent urease activity of 0.4470.01 mmol min À 1 mg protein À 1 and restored growth on urea as sole nitrogen source. Functional expression of the Nic1 transporter was essential for growth on urea at low Ni 2 þ concentrations. The maximum specific growth rates of the engineered strain on urea and ammonium were lower than those of a DUR1,2 reference strain. In glucose-limited chemostat cultures with urea as nitrogen source, the engineered strain exhibited an increased release of ammonia and reduced nitrogen content of the biomass. Our results indicate a new strategy for improving yeast-based production of nitrogen-containing chemicals and demonstrate that Ni 2 þ -dependent enzymes can be functionally expressed in S. cerevisiae.

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Section: Expression Of a Ni-dependent Atp-independent Urease In S Cesupporting

confidence: 80%

Section: Expression Of a Ni-dependent Atp-independent Urease In S Cementioning

confidence: 90%

See 1 more Smart Citation

Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

Milne

Luttik

Cueto-Rojas

et al. 2015

Metabolic Engineering

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show abstract

“…These yeasts catabolize urea via a urea amidolyase instead of the urease of other higher fungi (39). Deleting the urea amidolyase gene, DUR1,2, in C. albicans leads to a reduction in virulence and, importantly, of the fungal burden in organs with high urea content (40).…”

Section: Discussionmentioning

confidence: 99%

Histidine Degradation via an Aminotransferase Increases the Nutritional Flexibility of Candida glabrata

et al. 2014

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e The ability to acquire nutrients during infections is an important attribute in microbial pathogenesis. Amino acids are a valuable source of nitrogen if they can be degraded by the infecting organism. In this work, we analyzed histidine utilization in the fungal pathogen of humans Candida glabrata. Hemiascomycete fungi, like C. glabrata or Saccharomyces cerevisiae, possess no gene coding for a histidine ammonia-lyase, which catalyzes the first step of a major histidine degradation pathway in most other organisms. We show that C. glabrata instead initializes histidine degradation via the aromatic amino acid aminotransferase Aro8. Although ARO8 is also present in S. cerevisiae and is induced by extracellular histidine, the yeast cannot use histidine as its sole nitrogen source, possibly due to growth inhibition by a downstream degradation product. Furthermore, C. glabrata relies only on Aro8 for phenylalanine and tryptophan utilization, since ARO8, but not its homologue ARO9, was transcriptionally activated in the presence of these amino acids. Accordingly, an ARO9 deletion had no effect on growth with aromatic amino acids. In contrast, in S. cerevisiae, ARO9 is strongly induced by tryptophan and is known to support growth on aromatic amino acids. Differences in the genomic structure of the ARO9 gene between C. glabrata and S. cerevisiae indicate a possible disruption in the regulatory upstream region. Thus, we show that, in contrast to S. cerevisiae, C. glabrata has adapted to use histidine as a sole source of nitrogen and that the aromatic amino acid aminotransferase Aro8, but not Aro9, is the enzyme required for this process.

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“…Like all hemiascomycetes (yeasts), C. albicans uses the energy requiring bifunctional enzyme urea amidolyase (Dur1,2p), whereas other higher fungi use the nickel-containing urease [13]. The enzyme urea amidolyase, encoded by DUR1,2 (Degradation of URea), was first characterized in the yeast Candida utilis [14].…”

Section: Introductionmentioning

confidence: 99%

Urea Amidolyase (DUR1,2) Contributes to Virulence and Kidney Pathogenesis of Candida albicans

et al. 2012

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The intracellular enzyme urea amidolyase (Dur1,2p) enables C. albicans to utilize urea as a sole nitrogen source. Because deletion of the DUR1,2 gene reduces survival of C. albicans co-cultured with a murine macrophage cell line, we investigated the role of Dur1,2p in pathogenesis using a mouse model of disseminated candidiasis. A dur1,2Δ/dur1,2Δ strain was significantly less virulent than the wild-type strain, showing significantly higher survival rate, better renal function, and decreased and less sustained fungal colonization in kidney and brain. Complementation of the mutant restored virulence. DUR1,2 deletion resulted in a milder host inflammatory reaction. Immunohistochemistry, flow cytometry, and magnetic resonance imaging showed decreased phagocytic infiltration into infected kidneys. Systemic cytokine levels of wild-type mice infected with the dur1,2 mutant showed a more balanced systemic pro-inflammatory cytokine response. Host gene expression and protein analysis in infected kidneys revealed parallel changes in the local immune response. Significant differences were observed in the kidney IL-1 inflammatory pathway, IL-15 signaling, MAP kinase signaling, and the alternative complement pathway. We conclude that Dur1,2p is important for kidney colonization during disseminated candidiasis and contributes to an unbalanced host inflammatory response and subsequent renal failure. Therefore, this Candida-specific enzyme may represent a useful drug target to protect the host from kidney damage associated with disseminated candidiasis.

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Evolutionary aspects of urea utilization by fungi

Cited by 43 publications

References 26 publications

Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

Histidine Degradation via an Aminotransferase Increases the Nutritional Flexibility of Candida glabrata

Urea Amidolyase (DUR1,2) Contributes to Virulence and Kidney Pathogenesis of Candida albicans

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