<i>ALT1</i>-encoded alanine aminotransferase plays a central role in the metabolism of alanine in<i>Saccharomyces cerevisiae</i>

García-Campusano, Florencia García-CampusanoF.; Anaya, Víctor-Hugo AnayaV.-H.; Robledo-Arratia, Luis; Quezada, Héctor; Hernández-Barrios, Hugo; Riego, Lina; González, Alicia

doi:10.1139/w08-150

Cited by 28 publications

(25 citation statements)

References 25 publications

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“…Enhanced growth of Δ fmp12 cells was almost fully suppressed by a single disruption of the PUT1 (encoding proline dehydrogenase), PUT2 (encoding P5C dehydrogenase), ALT1 (encoding mitochondrial alanine transaminase) 14, or KGD1 (encoding E1 component of the mitochondrial α-ketoglutarate dehydrogenase (KGDH) complex) 15 gene, but not affected by single or combined deletion of the GDH2 (encoding NAD + -dependent glutamate dehydrogenase) and GDH1/3 (encoding NADP + -dependent glutamate dehydrogenases) genes involved in deamination of glutamate and its reverse reaction, respectively 51617, or by the GAD1 (encoding glutamate decarboxylase) gene, whose product catalyzes the first step of the γ-aminobutyrate (GABA) shunt 18 that bypasses the TCA cycle. Similar results were obtained on SD-N-C+Glu medium, although disruption of the PUT1 or PUT2 gene did not abolish the growth of Δ fmp12 cells.…”

Section: Resultsmentioning

confidence: 99%

Putative mitochondrial α-ketoglutarate-dependent dioxygenase Fmp12 controls utilization of proline as an energy source in Saccharomyces cerevisiae

Nishida¹,

Watanabe²,

Takagi³

2016

Microb Cell

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The amino acid proline functions as a nitrogen source and as a stress protectant in the yeast Saccharomyces cerevisiae. However, utilization of proline as a carbon source in S. cerevisiae cells has not been studied yet. In the process of study on the physiological roles of the found-in-mitochondrial-proteome (FMP) genes in proline metabolism, we found that Δfmp12 cells could grow better than wild-type cells on agar plate medium containing proline as the sole nitrogen and carbon sources. In contrast, overexpression of FMP12 negatively affected cell growth under the same condition. The Fmp12 protein was localized in the mitochondria and was constitutively expressed. Deletion of the genes that encode mitochondrial enzymes, such as proline dehydrogenase (PUT1), Δ1-pyrroline-5-carboxylate dehydrogenase (PUT2), alanine transaminase (ALT1), and α-ketoglutarate dehydrogenase subunit (KGD1), abolished the enhanced cell growth in Δfmp12. These results provided the first evidence that proline can be utilized as a carbon source via the mitochondrial proline metabolic pathway and the subsequent tricarboxylic acid (TCA) cycle in S. cerevisiae. The function of Fmp12, which has a similarity with α-ketoglutarate-dependent dioxygenases of the yeast Candida species and human, might inhibit cell growth by skipping the ATP production step of the TCA cycle.

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Section: Resultsmentioning

confidence: 99%

Putative mitochondrial α-ketoglutarate-dependent dioxygenase Fmp12 controls utilization of proline as an energy source in Saccharomyces cerevisiae

Nishida¹,

Watanabe²,

Takagi³

2016

Microb Cell

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“…Bolded arrows indicate pathway up-regulation whereas dashed arrows indicated pathway down-regulation relative to the wild-type strain. Potential alternative sources of glutamate are indicated by light gray arrows (Brandriss and Magasanik, 1980; Eden et al ., 1996; García-Campusano et al ., 2009; Iraqui et al ., 1998; Kispal et al ., 1996; Morin et al ., 1992; Urrestarazu et al ., 1998). Abbreviations: AATs = amino acid transaminases; ArAAs = aromatic amino acids; BCAAs = branched-chain amino acids; BCKA = branched chain keto acids.…”

Section: Figurementioning

confidence: 99%

Differential contribution of the proline and glutamine pathways to glutamate biosynthesis and nitrogen assimilation in yeast lacking glutamate dehydrogenase

Sieg

Trotter

2014

Microbiological Research

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In Saccharomyces cerevisiae, the glutamate dehydrogenase (GDH) enzymes play a pivotal role in glutamate biosynthesis and nitrogen assimilation. It has been proposed that, in GDH-deficient yeast, either the proline utilization (PUT) or the glutamine synthetase-glutamate synthase (GS/GOGAT) pathway serves as the alternative pathway for glutamate production and nitrogen assimilation to the exclusion of the other. Using a gdh-null mutant (gdh1Δ2Δ3Δ), this ambiguity was addressed using a combination of growth studies and pathway-specific enzyme assays on a variety of nitrogen sources (ammonia, glutamine, proline and urea). The GDH-null mutant was viable on all nitrogen sources tested, confirming that alternate pathways for nitrogen assimilation exist in the gdh-null strain. Enzyme assays point to GS/GOGAT as the primary alternative pathway on the preferred nitrogen sources ammonia and glutamine, whereas growth on proline required both the PUT and GS/GOGAT pathways. In contrast, growth on glucose-urea media elicited a decrease in GOGAT activity along with an increase in activity of the PUT pathway specific enzyme Δ1-pyrroline-5-carboxylate dehydrogenase (P5CDH). Together, these results suggest the alternative pathway for nitrogen assimilation in strains lacking the preferred GDH-dependent route is nitrogen source dependent and that neither GS/GOGAT nor PUT serves as the sole compensatory pathway.

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“…Especially for ALT2, its physiological role in vivo is not clear, although it is translated in yeast cells. ALT2 was reported to have no alanine aminotransferase activity, whereas its homologous enzyme ALT1 does in vivo [40], [41]. For CTP1, earlier Southern blot experiments indicated only one copy of the CTP1 gene exists in the S. cerevisiae genome.…”

Section: Discussionmentioning

confidence: 99%

Identification of Novel Knockout Targets for Improving Terpenoids Biosynthesis in Saccharomyces cerevisiae

Sun

Meng

et al. 2014

PLoS ONE

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Many terpenoids have important pharmacological activity and commercial value; however, application of these terpenoids is often limited by problems associated with the production of sufficient amounts of these molecules. The use of Saccharomyces cerevisiae (S. cerevisiae) for the production of heterologous terpenoids has achieved some success. The objective of this study was to identify S. cerevisiae knockout targets for improving the synthesis of heterologous terpeniods. On the basis of computational analysis of the S. cerevisiae metabolic network, we identified the knockout sites with the potential to promote terpenoid production and the corresponding single mutant was constructed by molecular manipulations. The growth rates of these strains were measured and the results indicated that the gene deletion had no adverse effects. Using the expression of amorphadiene biosynthesis as a testing model, the gene deletion was assessed for its effect on the production of exogenous terpenoids. The results showed that the dysfunction of most genes led to increased production of amorphadiene. The yield of amorphadiene produced by most single mutants was 8–10-fold greater compared to the wild type, indicating that the knockout sites can be engineered to promote the synthesis of exogenous terpenoids.

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ALT1-encoded alanine aminotransferase plays a central role in the metabolism of alanine inSaccharomyces cerevisiae

Cited by 28 publications

References 25 publications

Putative mitochondrial α-ketoglutarate-dependent dioxygenase Fmp12 controls utilization of proline as an energy source in Saccharomyces cerevisiae

Putative mitochondrial α-ketoglutarate-dependent dioxygenase Fmp12 controls utilization of proline as an energy source in Saccharomyces cerevisiae

Differential contribution of the proline and glutamine pathways to glutamate biosynthesis and nitrogen assimilation in yeast lacking glutamate dehydrogenase

Identification of Novel Knockout Targets for Improving Terpenoids Biosynthesis in Saccharomyces cerevisiae

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