Sequestration of arginine by polyphosphate in vacuoles of yeast (Saccharomyces cerevisiae)

Dürr, Mathias; Urech, Konrad; Böller, Th.; Wiemken, Andres; Schwencke, Jaime; Nagy, M

doi:10.1007/bf00689982

Cited by 108 publications

(43 citation statements)

References 20 publications

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“…Inasmuch as scPPX2 and scPPX3 are vacuolar activities, but only scPPX2 depends on proteolytic activation, it was uncertain whether scPPX3 is a distinctive enzyme or the residual activity of an unprocessed scPPX2. The two activities were therefore compared in vacuolar preparations with respect to the influence of arginine which is accumulated in the vacuole, where it acts as a counterion for polyP (9). With increasing arginine, the activity of scPPX3 increased, whereas the scPPX2 activity displayed a different pattern (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To determine whether the accumulation of polyP and its chain length are affected by reduced polyPase activity, polyP levels and size were determined by using 32 P in SDC medium with 7.35 mM phosphate. The extracts were obtained from acid-insoluble material after treatment with organic solvents and precipitation with Ba 2ϩ , a procedure which yields up to a 96% recovery of [ 32 P]polyP 750 added to unlabeled cells; acid-soluble polyP (polyP [3][4][5][6][7][8][9][10] ) is not recovered in this procedure. PolyP, measured by the release of P i after treatment with an excess of rPPX1, showed a slow accumulation during growth (Fig.…”

Section: Resultsmentioning

confidence: 99%

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The gene for a major exopolyphosphatase of Saccharomyces cerevisiae

1995

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) has been isolated from a genomic library. The gene, located at 57 kbp from the end of the right arm of chromosome VIII, encodes a protein of 396 amino acids. Overexpression in Escherichia coli allowed the ready purification of a recombinant form of the enzyme. Disruption of the gene did not affect the growth rate of S. cerevisiae. Lysates from the mutants displayed considerably lower exopolyphosphatase activity than the wild type. The enzyme is located in the cytosol, whereas the vast accumulation of polyphosphate (polyP) of the yeast is in the vacuole. Disruption of PPX1 in strains with and without deficiencies in vacuolar proteases allowed the identification of exopolyphosphatase activity in the vacuole. This residual activity was strongly reduced in the absence of vacuolar proteases, indicating a dependence on proteolytic activation. A 50-fold-lower protease-independent activity could be distinguished from this protease-dependent activity by different patterns of expression during growth and activation by arginine. With regard to the levels of polyP in various mutants, those deficient in vacuolar ATPase retain less than 1% of the cellular polyP, a loss that is not offset by additional mutations that eliminate the cytosolic exopolyphosphatase and the vacuolar polyphosphatases dependent on vacuolar protease processing.Inorganic polyphosphates (polyP) are widespread in nature and are particularly abundant in Saccharomyces cerevisiae, accounting for nearly 40% of the total phosphate content (22). The polyP chains of S. cerevisiae vary between three and several thousand P i residues (19). Among the various yeast cell compartments, the bulk is in the vacuoles (38), with lesser amounts and smaller chains found in mitochondria (4); in Saccharomyces fragilis, polyP has also been found on the cell surface (37). The distinct locations and sizes of polyP in yeast cells indicate either different pathways of biosynthesis and degradation or a sophisticated sorting system. These different polyP pools might be involved in a variety of functions, such as energy storage, P i reservoir, metal chelator, buffer against alkali, and regulator. None of these functions for polyP have been established in S. cerevisiae, whereas in Escherichia coli, a requirement for polyP for survival in stationary phase has been demonstrated (8).Polyphosphatase (polyPase) activity in S. cerevisiae is demonstrably high: a cell extract releases up to 130 nmol of P i per min per mg of yeast protein (42). In contrast, biosynthetic activity is rather feeble in vitro. Polyphosphate kinase, the enzyme that can make polyP in a reversible reaction by condensing the terminal phosphate of ATP, catalyzes the transfer of only about 4 nmol of phosphate residues from polyP to ADP per min per mg of yeast protein and is even less active in the synthesis of polyP (12,34). In view of the high level of polyPase activity, effective mechanisms must exist to protect polyP from hydrolytic degradation. Also implied are functions for the polyPase in using polyP as a reservoir...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The gene for a major exopolyphosphatase of Saccharomyces cerevisiae

1995

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“…For instance, arginine represents the largest nitrogen storage reserve in yeast, accumulating in vast quantities inside vacuoles (45) where it counteracts negatively charged polyphosphate polymers (46,47). These are long inorganic phosphate polymers that constitute the main phosphate repository in yeast (48).…”

Section: Discussionmentioning

confidence: 99%

Arginine Transcriptional Response Does Not Require Inositol Phosphate Synthesis

Bosch

Saiardi

2012

Journal of Biological Chemistry

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Background:The involvement of inositol phosphates produced by the yeast inositol-polyphosphate multikinase, Arg82, in transcription is controversial. Results: Catalytically inactive Arg82 restores the regulation of arginine-dependent genes in an ARG82 knock-out. Conclusion: Inositol phosphates do not regulate arginine-dependent gene expression. Significance: Independently of its enzymatic activity Arg82 controls arginine-responsive genes.

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“…Strullu et al (1981a) also pointed out that the ratio of calcium to phosphorus in mycorrhizal granules was only one-sixth that in calcium triphosphate and so presumably only one-half of that needed to satisfy the charge on the polyphosphate polyanions. They suggested that the more basic amino acids, such as glutamine or arginine, might balance the residue of the electrical charge, as found in Saccharomyces cerevisiae by Durr et al (1979).…”

Section: Discussionmentioning

confidence: 99%

Formation of Metachromatic Granules Following Phosphate Uptake by Mycelial Hyphae of an Ectomycorrhizal Fungus

1984

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SUMMARYAbundant metachromatic granules, presumed to contain polyphosphate, were present in mycelial hyphae oiPaxillus involutus (Batsch. ex Fr.) Fr. grown in phosphate-rich medium, but were absent when the medium lacked phosphate. Phosphorus-starved hyphae placed into phosphate-containing solutions developed granules within 3 to 4 h. Most of this period was required for synthesis of granule materials, since phosphate uptake occurred within minutes. Both phosphate uptake and granule formation are active processes, affected by low temperature and respiratory inhibitiors. No special importance seemed to attach to the presence of any of the following cations: Ca2+; Mg^+i K+; or Na+; and two different isolates of P. involutus, one calcifuge and one calcicole, showed no differential effects regarding granule formation in the presence and absence of Ca^+ and K ey words: Ectomycorrhizal fungus, metachromatic granule, polyphosphate, phosphate uptake, calcium.

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Sequestration of arginine by polyphosphate in vacuoles of yeast (Saccharomyces cerevisiae)

Cited by 108 publications

References 20 publications

The gene for a major exopolyphosphatase of Saccharomyces cerevisiae

The gene for a major exopolyphosphatase of Saccharomyces cerevisiae

Arginine Transcriptional Response Does Not Require Inositol Phosphate Synthesis

Formation of Metachromatic Granules Following Phosphate Uptake by Mycelial Hyphae of an Ectomycorrhizal Fungus

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