GDH1 expression is regulated by GLN3, GCN4, and HAP4 under respiratory growth

Riego, Lina; Avendaño, Amaranta; DeLuna, Alexander; Rodríguez, E; González, Alicia

doi:10.1016/s0006-291x(02)00174-2

Cited by 45 publications

(69 citation statements)

References 31 publications

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“…Further analysis of the GDH1 expression profile showed that it was also determined by the action of the GLN3-encoded transcriptional activator. gln3D and hap2D mutants displayed similar expression levels, suggesting that both regulators could coordinately determine GDH1 transcription (Riego et al, 2002).…”

Section: Resultsmentioning

confidence: 97%

“…Mutants simultaneously affected in the GATAA (3) , GATAA (4) and GATAA (5) sites present in the GDH1 promoter (Riego et al, 2002) were constructed by transforming strain CLA-1 (MATa ure3 leu2 GLN3-myc 13 NAT) with a 3.2 kb fragment obtained by PCR amplification of the pCORE plasmid harbouring the kanMX4, KlURA3 CORE module, using the pCORE/gata Gdh1 Fw and pCORE/gata Gdh1 Rv oligonucleotides described in Table 3 ( Storici & Resnick, 2003). Transformation was carried out following the protocol described by Longtine et al (1998).…”

Section: Methodsmentioning

confidence: 99%

“…Correct insertion was verified by PCR amplification. One of the transformants was retransformed with IROs (integrative recombinant oligonucleotides) harbouring mutagenized GATAA (3) , GATAA (4) (GcaAAG) sequences (IRO 1-2 gln3 Cm Fw and IRO 1-2 gln3 Cm Rv; Table 3) (Riego et al, 2002). Correct insertion was confirmed by sequencing.…”

Section: Methodsmentioning

confidence: 99%

“…To analyse whether the GATAAG(C) cis-acting elements present in the GDH1 promoter (Riego et al, 2002) played a role in the Hap2-3-5-Gln3-mediated response, a mutant impaired in the GATAAG(C) (3, 4 and 5) sites was constructed and GDH1 transcriptional activation and Gln3 promoter occupancy were analysed.…”

Section: Gln3 Binding To the Gdh1 Promoter Is Hap2-dependentmentioning

confidence: 99%

“…Thus, under repressive nitrogen conditions, the role of Gln3 and Gat1 as activators is hindered. However, previous work from our laboratory showed that expression of genes pertaining to nitrogen assimilation pathways, such as GLT1, encoding glutamate synthase, and GDH1, encoding one of the two isoforms of glutamate dehydrogenase, was Gln3-dependent under repressive nitrogen conditions, suggesting that Gln3 could determine gene expression even under repressive conditions (Riego et al, 2002;Valenzuela et al, 1998). Most interesting was the finding that GDH1 expression was induced by the HAP complex, whose activator role had been considered to exclusively determine transcription of genes involved in the diauxic shift, mitochondrial biogenesis and energy metabolism, allowing adaptation of yeast from fermentative to respiratory growth (Dang et al, 1994).…”

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confidence: 91%

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Hap2-3-5-Gln3 determine transcriptional activation of GDH1 and ASN1 under repressive nitrogen conditions in the yeast Saccharomyces cerevisiae

et al. 2011

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The transcriptional activation response relies on a repertoire of transcriptional activators, which decipher regulatory information through their specific binding to cognate sequences, and their capacity to selectively recruit the components that constitute a given transcriptional complex. We have addressed the possibility of achieving novel transcriptional responses by the construction of a new transcriptional regulator -the Hap2-3-5-Gln3 hybrid modulator -harbouring the HAP complex polypeptides that constitute the DNA-binding domain (Hap2-3-5) and the Gln3 activation domain, which usually act in an uncombined fashion. The results presented in this paper show that transcriptional activation of GDH1 and ASN1 under repressive nitrogen conditions is achieved through the action of the novel Hap2-3-5-Gln3 transcriptional regulator. We propose that the combination of the Hap DNA-binding and Gln3 activation domains results in a hybrid modulator that elicits a novel transcriptional response not evoked when these modulators act independently. INTRODUCTIONThe response of Saccharomyces cerevisiae to nutrient limitation is a model whose study has amply contributed to our knowledge of the mechanisms determining transcriptional activation (Zaman et al., 2008). Nutrient limitation results in a complex adaptation of the physiology of yeast cells, which allows them to redefine their transcription programme. This leads to the activation of particular sets of genes, whose products are needed to evoke an appropriate physiological response. Transcriptional activators are composed of a DNA-binding domain, which targets these proteins to specialized binding sites, and an activation domain that mediates transcription initiation; these two domains can be contained in single or different polypeptides (Zaman et al., 2008).When yeast cells are provided with poor nitrogen sources, such as proline, genes coding for enzymes involved in the catabolism of these compounds are highly expressed. Conversely, in the presence of high-quality nitrogen sources such as glutamine, a decrease in the levels of catabolic enzymes is observed. The reduced expression of the genes coding for enzymes involved in the utilization of poor nitrogen sources is brought about through the action of a regulatory system known as nitrogen catabolite repression (NCR) (Blinder & Magasanik, 1995;Coffman et al., 1995Coffman et al., , 1996Courchesne & Magasanik, 1988;Minehart & Magasanik, 1991) NCR operates through the action of two transcriptional activators, Gln3 and Nil1/ Gat1, and two repressors, Dal80 and Gzf3/Deh1/Nil2 (Minehart & Magasanik, 1991;Stanbrough et al., 1995;Svetlov & Cooper, 1998;Magasanik & Kaiser, 2002), whose expression is regulated by nitrogen levels. In the presence of repressive nitrogen sources, the TOR signalling pathway promotes the association of the GATA transcription factors Gln3 and Gat1 with the cytoplasmic protein Ure2, thus retaining Gln3 and Gat1 in the cytoplasm (Beck & Hall, 1999;Cardenas et al., 1999;Hardwick et al., 1999). Dissociation o...

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Section: Resultsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Gln3 Binding To the Gdh1 Promoter Is Hap2-dependentmentioning

confidence: 99%

mentioning

confidence: 91%

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Hap2-3-5-Gln3 determine transcriptional activation of GDH1 and ASN1 under repressive nitrogen conditions in the yeast Saccharomyces cerevisiae

et al. 2011

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Diversification of the kinetic properties of yeast NADP‐glutamate‐dehydrogenase isozymes proceeds independently of their evolutionary origin

et al. 2016

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In the yeast Saccharomyces cerevisiae, the ScGDH1 and ScGDH3 encoded glutamate dehydrogenases (NADP‐GDHs) catalyze the synthesis of glutamate from ammonium and α‐ketoglutarate (α‐KG). Previous kinetic characterization showed that these enzymes displayed different allosteric properties and respectively high or low rate of α‐KG utilization. Accordingly, the coordinated action of ScGdh1 and ScGdh3, regulated balanced α‐KG utilization for glutamate biosynthesis under either fermentative or respiratory conditions, safeguarding energy provision. Here, we have addressed the question of whether there is a correlation between the regulation and kinetic properties of the NADP‐GDH isozymes present in S. cerevisiae (ScGdh1 and ScGdh3), Kluyveromyces lactis (KlGdh1), and Lachancea kluyveri (LkGdh1) and their evolutionary history. Our results show that the kinetic properties of K. lactis and L. kluyveri single NADP‐GDHs are respectively similar to either ScGDH3 or ScGDH1, which arose from the whole genome duplication event of the S. cerevisiae lineage, although, KlGDH1 and LkGDH1 originated from a GDH clade, through an ancient interspecies hybridization event that preceded the divergence between the Saccharomyces clade and the one containing the genera Kluyveromyces, Lachancea, and Eremothecium. Thus, the kinetic properties which determine the NADP‐GDHs capacity to utilize α‐KG and synthesize glutamate do not correlate with their evolutionary origin.

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Glutamate dehydrogenases in the oleaginous yeast Yarrowia lipolytica

Trotter

Juco

et al. 2019

Yeast

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Glutamate dehydrogenases (GDHs) are fundamental to cellular nitrogen and energy balance. Yet little is known about these enzymes in the oleaginous yeast Yarrowia lipolytica. The YALI0F17820g and YALI0E09603g genes, encoding potential GDH enzymes in this organism, were examined. Heterologous expression in gdh‐null Saccharomyces cerevisiae and examination of Y. lipolytica strains carrying gene deletions demonstrate that YALI0F17820g (ylGDH1) encodes a NADP‐dependent GDH whereas YALI0E09603g (ylGDH2) encodes a NAD‐dependent GDH enzyme. The activity encoded by these two genes accounts for all measurable GDH activity in Y. lipolytica. Levels of the two enzyme activities are comparable during logarithmic growth on rich medium, but the NADP‐ylGDH1p enzyme activity is most highly expressed in stationary and nitrogen starved cells by threefold to 12‐fold. Replacement of ammonia with glutamate causes a decrease in NADP‐ylGdh1p activity, whereas NAD‐ylGdh2p activity is increased. When glutamate is both carbon and nitrogen sources, the activity of NAD‐ylGDH2p becomes dominant up to 18‐fold compared with that of NADP‐ylGDH1p. Gene deletion followed by growth on different carbon and nitrogen sources shows that NADP‐ylGdh1p is required for efficient nitrogen assimilation whereas NAD‐ylGdh2p plays a role in nitrogen and carbon utilization from glutamate. Overexpression experiments demonstrate that ylGDH1 and ylGDH2 are not interchangeable. These studies provide a vital basis for future consideration of how these enzymes function to facilitate energy and nitrogen homeostasis in Y. lipolytica.

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GDH1 expression is regulated by GLN3, GCN4, and HAP4 under respiratory growth

Cited by 45 publications

References 31 publications

Hap2-3-5-Gln3 determine transcriptional activation of GDH1 and ASN1 under repressive nitrogen conditions in the yeast Saccharomyces cerevisiae

Hap2-3-5-Gln3 determine transcriptional activation of GDH1 and ASN1 under repressive nitrogen conditions in the yeast Saccharomyces cerevisiae

Diversification of the kinetic properties of yeast NADP‐glutamate‐dehydrogenase isozymes proceeds independently of their evolutionary origin

Glutamate dehydrogenases in the oleaginous yeast Yarrowia lipolytica

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