Isolation and characterization of the gene encoding phosphoenolpyruvate carboxykinase from <i>Saccharomyces cerevisiae</i>

Valdes-Hevia, Marta D; Guerra, R. de la; Gancedo, Carlos

doi:10.1016/0014-5793(89)81682-5

Cited by 46 publications

(26 citation statements)

References 21 publications

(5 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A conceptual translation of the annotation fragment resulted in a 607 AA fragment having homology to PCK1 of S. cerevisiae (65% identity to NP013023). PCK1 encodes the enzyme phosphoenolpyruvate carboxykinase, which catalyzes the only irreversible step in gluconeogenesis and is a major regulatory checkpoint for the control of gluconeogenesis (42,43). Targeted knockout strains of PCK1 were constructed and confirmed by Southern blots digested with BamHI/KpnI and hybridized with a PCK1 fragment ( Figure 6B; WT: 8.0, 6.0, 1.1 kb; Δpck1: 8.0, 6.0 kb).…”

Section: Figurementioning

confidence: 99%

The DEAD-box RNA helicase Vad1 regulates multiple virulence-associated genes in Cryptococcus neoformans

Panepinto¹,

Liu²,

Ramos³

et al. 2005

J. Clin. Invest.

101

View full text Add to dashboard Cite

The study of fungal regulatory networks is essential to the understanding of how these pathogens respond to host environmental signals with effective virulence-associated traits. In this study, a virulence-associated DEAD-box RNA helicase-encoding gene (VAD1) was isolated from a mutant defective in the virulence factor laccase. A Δvad1 mutant exhibited a profound reduction in virulence in a mouse model that was restored after reconstitution with WT VAD1. Loss of VAD1 resulted in upregulation of NOT1, a gene encoding a global repressor of transcription. NOT1 was found to act as an intermediary transcriptional repressor of laccase. Vad1 was located within macromolecular complexes that formed cytoplasmic granular bodies in mature cells and during infection of mouse brain. In addition, VAD1 was shown by in situ hybridization to be expressed in the brain of an AIDS patient coinfected with C. neoformans. To understand the role of VAD1 in virulence, a functional genomics approach was used to identify 3 additional virulence determinants dependent on VAD1: PCK1, TUF1, and MPF3, involved in gluconeogenesis, mitochondrial protein synthesis, and cell wall integrity, respectively. These data show that fungal virulence-associated genes are coordinately regulated and that an analysis of such transcriptomes allows for the identification of important new genes involved in the normal growth and virulence of fungal pathogens.

show abstract

Section: Figurementioning

confidence: 99%

The DEAD-box RNA helicase Vad1 regulates multiple virulence-associated genes in Cryptococcus neoformans

Panepinto¹,

Liu²,

Ramos³

et al. 2005

J. Clin. Invest.

101

View full text Add to dashboard Cite

show abstract

“…SFC1 is coregulated with ICL1 and MLS1 and encodes a putative mitochondrial succinate-fumarate antiporter that may be involved in shuttling intermediates of the glyoxylate pathway into the appropriate subcellular locations (Fernandez et al, 1994;Lorenz and Fink, 2001). PCK1 and FBP1 encode phosphoenolpyruvate carboxykinase and fructose 1,6-bisphosphatase, respectively, which catalyze the two irreversible reactions of gluconeogenesis (Entian et al, 1988;Valdes-Hevia et al, 1989). Mutants lacking any of these five genes grew poorly using acetate as the sole carbon source, and all of these mutants but fbp1⌬ were able to grow well using glycerol as the sole carbon source (our unpublished data), demonstrating that these strains are competent for respiration but defective in the conversion of acetate to glucose.…”

Section: Mutants Of the Glyoxylate Pathway Form Nsds On Acetate Solidmentioning

confidence: 99%

Regulation of Spindle Pole Function by an Intermediary Metabolite

Nickas

Diamond

Yang

et al. 2004

MBoC

View full text Add to dashboard Cite

Spore formation in the yeast Saccharomyces cerevisiae depends on a modification of spindle pole bodies (SPBs) at the onset of meiosis II that allows them to promote de novo membrane formation. Depletion of the environmental carbon source during sporulation results in modification of only one SPB from each meiosis II spindle and formation of a two-spored ascus, called a nonsister dyad (NSD). We have found that mutants impaired in the glyoxylate pathway, which is required for the conversion of acetate to glucose, make NSDs when acetate is the primary carbon source. Wild-type cells make NSDs when the carbon source is glycerol, which is converted to glucose independently of the glyoxylate pathway. During NSD formation in glycerol, only the two SPBs created at the meiosis I/II transition ("daughters") are modified. In these conditions, the SPB components Mpc70p and Spo74p are not recruited to mother SPBs. Moreover, cooverexpression of Mpc70p and Spo74p suppresses NSD formation in glycerol. Our findings indicate that flux through the glyoxylate pathway during sporulation regulates modification of mother SPBs via recruitment of Mpc70p and Spo74p. These results define a cellular response in which the accumulation of an intermediary metabolite serves as a measure of biosynthetic capacity to regulate the number of daughter cells formed.

show abstract

“…Sci. USA 90 (1993) inactivation does not operate during prolonged growth on glucose, or at least that its capacity is limited (13,15).…”

Section: Estimation Of Fbpase Activity In the Transformed Strainsmentioning

confidence: 99%

Futile cycles in Saccharomyces cerevisiae strains expressing the gluconeogenic enzymes during growth on glucose.

Navas

Cerdán

Gancedo

1993

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The systems which control the levels of the gluconeogenic enzymes in Saccharomyces cerevisiae have been bypassed to ascertain their physiological significance. The coding regions of the genes FBPI and PCKI, which encode fructose-1,6-bisphosphatase and phosphoenolpyruvate carboxykinase, have been put under the control of the promoter of ADCi (alcohol dehydrogenase I), a gene not repressed by glucose, and introduced into yeast in multicopy plasmids. The transformed yeast cells show high levels of the gluconeogenic enzymes during growth on glucose. Generation time and growth yield of yeast expressing either fructose-1,6-bisphosphatase or phosphoenolpyruvate carboxykinase are not significantly different from those of the wild-type strain. For a strain expressing both enzymes the increase in generation time is about 20% and the decrease in growth yield around 30%. The concentration of ATP is about 1.5 mM in the growing cells of the different strains. The extent of in vivo cycling was measured by 13C NMR in cell-free extracts from yeast growing on [6-'3C]glucose. Cycling between fructose-6-phosphate and fructose-1,6-bisphosphate is <2%, most likely due to the very strong inhibition of fructose-1,6-bisphosphatase by fructose 2,6-bisphosphate. Cycling between phosphoenolpyruvate and pyruvate is low, but a precise figure could not be obtained due to poor equilibration of label between carbons 2 and 3 of oxaloacetate.activator fructose 1,6-bisphosphate (5). In contrast, the amount of the gluconeogenic enzymes FbPase and PEPCK is tightly regulated. Their synthesis is strongly repressed by glucose (6, 7) and they are proteolytically degraded when glucose is added to a derepressed yeast (8-10), a process known as catabolite inactivation (11). In addition, yeast FbPase is inhibited by Fru-2,6-P2 (12) and by AMP (6). The existence of a multilayered regulation of the gluconeogenic enzymes through catabolite repression, catabolite inactivation, and, at least for FbPase, allosteric control suggests that the different mechanisms have been selected to avoid the operation offutile cycles in yeast growing on glucose. A yeast expressing FbPase under the control of the PH05 promoter is able to grow on glucose (13), but no attempts have been made to determine whether cycling takes place when some of the controls on FbPase or PEPCK are removed and how this cycling affects the growth of the yeast.To investigate the physiological relevance of catabolite repression and catabolite inactivation of the gluconeogenic enzymes, we have put the genes encoding FbPase and PEPCK under the control of a promoter not repressed by glucose. We found that the expression of the gluconeogenic enzymes under glycolytic conditions has no striking consequences on yeast survival.Saccharomyces cerevisiae can grow on sugars, metabolized through the glycolytic pathway, or on 2-C or 3-C compounds, which require gluconeogenesis. Gluconeogenesis proceeds by a reversal of glycolysis except for the reactions catalyzed by phosphofructokinase and pyruvate kinase. T...

show abstract

Isolation and characterization of the gene encoding phosphoenolpyruvate carboxykinase from Saccharomyces cerevisiae

Cited by 46 publications

References 21 publications

The DEAD-box RNA helicase Vad1 regulates multiple virulence-associated genes in Cryptococcus neoformans

The DEAD-box RNA helicase Vad1 regulates multiple virulence-associated genes in Cryptococcus neoformans

Regulation of Spindle Pole Function by an Intermediary Metabolite

Futile cycles in Saccharomyces cerevisiae strains expressing the gluconeogenic enzymes during growth on glucose.

Contact Info

Product

Resources

About