Correction: Glutamate dehydrogenase (Gdh2)-dependent alkalization is dispensable for escape from macrophages and virulence of Candida albicans

Silao, Fitz Gerald S.; Ryman, Kicki; Jiang, Tong; Ward, Matthew; Hansmann, Nicolas; Molenaar, Chris; Liu, Ningning; Chen, Changbin; Ljungdahl, Per O.

doi:10.1371/journal.ppat.1009877

Cited by 2 publications

(14 citation statements)

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“…Consistently, recent work in our group and others have shown that enzymes of the proline catabolic pathway (e.g., Put1 and Put2) can be induced in the presence of preferred nitrogen sources (e.g., ammonium or amino acids) [59][60][61] and even in a strain lacking Gln3 and Gat1 [59]. In addition, the gene encoding glutamate dehydrogenase (GDH2), a key player in central nitrogen metabolism, is robustly expressed when there is an abundance of preferred nitrogen sources such as ammonium and amino acids, indicating that its expression is independent of NCR [61]. This latter finding is in striking contrast to S. cerevisiae, with its GDH2 subject to tight NCR control (reviewed in [47]).…”

Section: Figure 3 Schematic Diagram Of Nitrogen Catabolite Repression (Ncr) In Yeast (Adapted Fromsupporting

confidence: 86%

“…Certain amino acids traditionally classified as poor, such as proline in S. cerevisiae [47,58], are readily utilized by C. albicans lacking Gln3 and Gat1, clearly indicating that proline utilization is not subject to tight NCR control [56]. Consistently, recent work in our group and others have shown that enzymes of the proline catabolic pathway (e.g., Put1 and Put2) can be induced in the presence of preferred nitrogen sources (e.g., ammonium or amino acids) [59][60][61] and even in a strain lacking Gln3 and Gat1 [59]. In addition, the gene encoding glutamate dehydrogenase (GDH2), a key player in central nitrogen metabolism, is robustly expressed when there is an abundance of preferred nitrogen sources such as ammonium and amino acids, indicating that its expression is independent of NCR [61].…”

Section: Figure 3 Schematic Diagram Of Nitrogen Catabolite Repression (Ncr) In Yeast (Adapted Fromsupporting

confidence: 68%

“…The bulk of ammonia produced by C. albicans during growth on amino acids as a sole nitrogen and carbon source comes from the reaction catalyzed by Gdh2 [61]. Contrary to the initial publication, now corrected [61], Gdh2 in C. albicans is clearly cytoplasmic. The correct assignment of Gdh2 as a cytoplasmic component is key to understanding its role in cellular energy production, as the reaction reduces NAD + , forming NADH.…”

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 92%

“…Glutamate is enzymatically converted to α-KG via oxidative deamination, catalyzed by NAD + -dependent glutamate dehydrogenase (Gdh2; EC 1.4.1.2), yielding ammonia (NH 3 ) and reduced NADH [61,93]. At a physiologically relevant pH, ammonia is protonated to ammonium (NH 4 + ).…”

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 99%

“…In mammalian cells, glutamate dehydrogenase is localized in the mitochondria, whereas in S. cerevisiae, there is a lack of consensus regarding its localization; Gdh2 has been reported to be a mitochondrial component [94,95] and alternatively a cytosolic component [96][97][98]. The bulk of ammonia produced by C. albicans during growth on amino acids as a sole nitrogen and carbon source comes from the reaction catalyzed by Gdh2 [61]. Contrary to the initial publication, now corrected [61], Gdh2 in C. albicans is clearly cytoplasmic.…”

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 99%

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Amino Acid Sensing and Assimilation by the Fungal Pathogen Candida albicans in the Human Host

Silao

Ljungdahl

2021

Pathogens

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Nutrient uptake is essential for cellular life and the capacity to perceive extracellular nutrients is critical for coordinating their uptake and metabolism. Commensal fungal pathogens, e.g., Candida albicans, have evolved in close association with human hosts and are well-adapted to using diverse nutrients found in discrete host niches. Human cells that cannot synthesize all amino acids require the uptake of the “essential amino acids” to remain viable. Consistently, high levels of amino acids circulate in the blood. Host proteins are rich sources of amino acids but their use depends on proteases to cleave them into smaller peptides and free amino acids. C. albicans responds to extracellular amino acids by pleiotropically enhancing their uptake and derive energy from their catabolism to power opportunistic virulent growth. Studies using Saccharomyces cerevisiae have established paradigms to understand metabolic processes in C. albicans; however, fundamental differences exist. The advent of CRISPR/Cas9-based methods facilitate genetic analysis in C. albicans, and state-of-the-art molecular biological techniques are being applied to directly examine growth requirements in vivo and in situ in infected hosts. The combination of divergent approaches can illuminate the biological roles of individual cellular components. Here we discuss recent findings regarding nutrient sensing with a focus on amino acid uptake and metabolism, processes that underlie the virulence of C. albicans.

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Section: Figure 3 Schematic Diagram Of Nitrogen Catabolite Repression (Ncr) In Yeast (Adapted Fromsupporting

confidence: 86%

Section: Figure 3 Schematic Diagram Of Nitrogen Catabolite Repression (Ncr) In Yeast (Adapted Fromsupporting

confidence: 68%

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 92%

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 99%

Section: Amino Acid Metabolism In C Albicansmentioning

confidence: 99%

See 3 more Smart Citations

Amino Acid Sensing and Assimilation by the Fungal Pathogen Candida albicans in the Human Host

Silao

Ljungdahl

2021

Pathogens

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Correction: Correction: Glutamate dehydrogenase (Gdh2)-dependent alkalization is dispensable for escape from macrophages and virulence of Candida albicans

2021

PLoS Pathog

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Supporting informationS1 Fig. CRISPR/Cas9-mediated gene inactivation of GDH2 and construction of a GDH2-GFP reporter strain. (A) A purified KpnI/SacI fragment from pFS108, harboring GDH2-specific sgRNA, and PCR generated repair template (RT) were introduced into wildtype strain SC5314 by electroporation. Nou R transformants were pre-screened in YNB+Arg medium containing the pH indicator bromocresol purple; the initial pH was 4.0. Three Nou R colonies were picked for further analysis. Clones #1 and #2 grew poorly and were unable to alkalinize the media; clone #3 grew and alkalinized the media. (B) Genomic DNA, isolated from the three clones, was used as template for PCR amplification of the targeted GDH2 locus; ddH 2 O was used as negative control. Restriction of the amplified �900 bp fragment by XhoI is diagnostic for successful mutagenesis (primers p5/p6; S2 Table ). Strains, clone #1 (CFG277) and clone #2 (CFG278), carry inactivated gdh2-/-alleles. (C) GDH2 is not essential but required for robust growth on glutamate or proline as sole nitrogen source. Five microliters of serially diluted wildtype (SC5314), gdh2-/-NAT R (CFG277), gdh2-/-NAT S (CFG279), and control (CFG182) cells were spotted on yeast peptone (YP), synthetic glutamate (SE) and synthetic proline (SP) media containing either 2% glucose (YPD, SED, SPD) or 1% glycerol (YPG, SEG, SPG) as carbon source. The plates were incubated for 48 h at 30 ˚C and photographed. (D) Fresh colonies of SC5314 (PLC005; WT), CFG279 (gdh2-/-), CASJ041 (cph1-/-efg1-/-) and CFG352 (cph1-/-efg1-/-gdh2-/-) were individually resuspended in YNB+CAA medium and incubated for 24 h at 37 ˚C. (E) The insertion of GFP in strain CFG273 (GDH2-GFP) was verified by PCR, the expected 1695 bp fragment was amplified using primers (p24/p25; S2 Table ); strain CAI4 served as untagged control (middle left panel). CFG273 was transformed with the CRISPR/Cas9 cassette to inactivate GDH2. Putative gdh2-/-clones were identified as described and verified by PCR-RD (p13/p6; S2 Table ) resulting in strain CFG400. Strains CFG273 (GDH2/GDH2-GFP) and CFG400 (gdh2/gdh2-GFP) were grown at a starting OD 600 � 2 in SE medium with 0.2% glucose and 1 mM proline (SED 0.2%+Pro) for 2 h. In contrast to CFG273, strain CFG400 failed to express Gdh2-GFP as assessed by immunoblot (middle right panel) and microscopy (lower panels; Scale bar = 5 μm), demonstrating the specificity of CRISPR/ Cas9.

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Correction: Glutamate dehydrogenase (Gdh2)-dependent alkalization is dispensable for escape from macrophages and virulence of Candida albicans

Cited by 2 publications

References 15 publications

Amino Acid Sensing and Assimilation by the Fungal Pathogen Candida albicans in the Human Host

Amino Acid Sensing and Assimilation by the Fungal Pathogen Candida albicans in the Human Host

Correction: Correction: Glutamate dehydrogenase (Gdh2)-dependent alkalization is dispensable for escape from macrophages and virulence of Candida albicans

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