Regulation of yeast fructose‐1,6‐bisphosphatase in strains containing multicopy plasmids coding for this enzyme

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

doi:10.1016/0014-5793(88)81004-4

Cited by 27 publications

(19 citation statements)

References 26 publications

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“…These results show that catabolite repression does not affect the levels of the YlFBP1 mRNA and that YlFbp is not subject to catabolite inactivation. [16]), no insertion (pDB20 [7]), the YlFBP1 gene under the control of the ScFBP1 promoter (pCL201), the ScFBP1 promoter (pCL202), the YlFBP1 gene under the control of the ScADH1 promoter (pCL203), or a similar plasmid with the YlFBP1 gene in the inverted orientation (pCL204). A purified transformant of each was streaked on plates with glucose or with glycerol plus ethanol as carbon sources.…”

Section: Resultsmentioning

confidence: 99%

“…Plasmid pCL201 is an episomal plasmid that carries a 600-bp SmaIEcoRV DNA fragment from the S. cerevisiae FBP1 (ScFBP1) promoter and the first 18 codons of ScFBP1 in frame with the YlFBP1 ORF in plasmid pDB20 (7) digested with SmaI and NotI; pCL202 is similar to pCL201 but carries only the ScFBP1 promoter fragment; pCL203 expresses the YlFBP1 ORF under the control of the ScADH1 promoter in the episomal plasmid pDB20 (7); and pCL204 is similar but with YlFBP1 in the inverted orientation. Other plasmids used were pRG6, an episomal plasmid bearing the S. cerevisiae FBP1 gene (16), and pINA237, which carries the YlLEU2 gene (23).…”

Section: Methodsmentioning

confidence: 99%

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The Gluconeogenic Enzyme Fructose-1,6-Bisphosphatase Is Dispensable for Growth of the Yeast Yarrowia lipolytica in Gluconeogenic Substrates

2008

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The genes encoding gluconeogenic enzymes in the nonconventional yeast Yarrowia lipolytica were found to be differentially regulated. The expression of Y. lipolytica FBP1 (YlFBP1) encoding the key enzyme fructose-1,6-bisphosphatase was not repressed by glucose in contrast with the situation in other yeasts; however, this sugar markedly repressed the expression of YlPCK1, encoding phosphoenolpyruvate carboxykinase, and YlICL1, encoding isocitrate lyase. We constructed Y. lipolytica strains with two different disrupted versions of YlFBP1 and found that they grew much slower than the wild type in gluconeogenic carbon sources but that growth was not abolished as happens in most microorganisms. We attribute this growth to the existence of an alternative phosphatase with a high K m (2.3 mM) for fructose-1,6-bisphosphate. The gene YlFBP1 restored fructose-1,6-bisphosphatase activity and growth in gluconeogenic carbon sources to a Saccharomyces cerevisiae fbp1 mutant, but the introduction of the FBP1 gene from S. cerevisiae in the Ylfbp1 mutant did not produce fructose-1,6-bisphosphatase activity or growth complementation. Subcellular fractionation revealed the presence of fructose-1,6-bisphosphatase both in the cytoplasm and in the nucleus.The growth of yeasts and other microorganisms in nonsugar carbon sources is dependent on gluconeogenesis. This pathway implicates specific gluconeogenic enzymes, such as fructose-1,6-bisphosphatase (Fbp) and phosphoenolpyruvate carboxykinase (Pck), that bypass the two physiologically irreversible steps in the glycolytic pathway, namely, phosphofructokinase and pyruvate kinase. In addition, under certain growth conditions, the enzymes from the glyoxylate cycle, isocitrate lyase (Icl) and malate synthase, are also required for the function of gluconeogenesis (Fig. 1). Growth in any nonsugar carbon source requires Fbp, which catalyzes the hydrolysis of fructose-1,6-P 2 to fructose-6-phosphate, and therefore, Fbp is a key enzyme in gluconeogenesis. The simultaneous function of specific gluconeogenic enzymes and their glycolytic counterparts would lead to futile cycles, i.e., reactions that waste ATP without yielding a net product (42). Presumably due to evolutionary pressure, a series of control mechanisms have been selected to minimize the functioning of those cycles. In the yeast Saccharomyces cerevisiae, the activities of phosphofructokinase and pyruvate kinase are controlled by a series of activators and inhibitors whose concentrations vary upon growth on sugars or nonsugar carbon sources (12,61,65). The activity of gluconeogenic enzymes is determined by the interplay of a variety of mechanisms. The transcription of the genes encoding them is subjected to catabolite repression, a complex regulatory phenomenon that involves many proteins (13,32,41). In addition, in S. cerevisiae, most of the gluconeogenic enzymes undergo catabolite inactivation (27, 48, 50), a proteolytic degradation that destroys them after glucose addition. Control by metabolites appears to be restricted to Fbp that ...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Gluconeogenic Enzyme Fructose-1,6-Bisphosphatase Is Dispensable for Growth of the Yeast Yarrowia lipolytica in Gluconeogenic Substrates

2008

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“…Catabolite inactivation of PEPCK, FbPase and malate dehydrogenase in yeast strains carrying multicopy or integrative plasmids with the PCK gene. Inactivation was performed as described [23]. Squares, PEPCK; circles, FbPase; triangles, malate dehydrogenase.…”

Section: Resultsmentioning

confidence: 99%

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

1989

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The yeast PCKl gene codmg for phosphoenolpy~vate carboxykmasc (PEPCK) was Isolated by functIona complementatlon of pckl strains from S cerevzsfae Only one copy of the gene was found per baplold yeast gcnome An RNA of about 2 kb which hybndlzed with a DNA probe mtemal to the PCKl gene was found only m cells growmg m non-fermentable carbon sources Yeast &rams carrymg multiple copies of the PCKl gene showed normal catabohte represslon of PEPCK except those carrying the shortest msertlon complementmg the mutation (2 2 kb) that presented an altered kmetlcs of derepresslon Catabohte mactlvatlon was decreased m strains transfomed with multtcopy plasmlds carrymg the PCKl gene Phosphoenolpyruvate carboxykmase, Gluconeogenesls, Catabohte repression, Catabohte mactlvatlon

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“…Again, a functional redundancy is observed in the PYC reaction, as two genes have been found (STUCKA et al, 1991). Interestingly, overproduction of FBPase in cells growing on glucose does not have a marked effect (DE LA GUERRA et al, 1988). Under these as well as under gluconeogenic growth conditions catabolic and anabolic enzymes are present simultaneously, indicating the need for allosteric mechanisms to avoid waste of energy (see Sect.…”

Section: Gluconeogenesismentioning

confidence: 97%

“…Aside from the predominant utilization of S. cerevisiae for fermentation processes, other yeasts are of potential industrial use, e.g., in the production of lipases for highly specific modifications of oil components (BJ~RKLING et al, 1991;DEL RIO et al, 1990), in the conversion of pentoses, the principal components of hemicelluloses (JEFFRIES, 1990), and in the use of killer strains in biotyping of pathogenic yeasts (POLONELLI et al, 1991). The use of baker's yeast as a reagent in the synthesis of various organic compounds has been reviewed recently (SERVI, 1990).…”

Section: "Classical" Applicationsmentioning

confidence: 99%

Yeasts

Heinisch¹,

Hollenberg²

2001

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Regulation of yeast fructose‐1,6‐bisphosphatase in strains containing multicopy plasmids coding for this enzyme

Cited by 27 publications

References 26 publications

The Gluconeogenic Enzyme Fructose-1,6-Bisphosphatase Is Dispensable for Growth of the Yeast Yarrowia lipolytica in Gluconeogenic Substrates

The Gluconeogenic Enzyme Fructose-1,6-Bisphosphatase Is Dispensable for Growth of the Yeast Yarrowia lipolytica in Gluconeogenic Substrates

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

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