The early stage of polyphosphate accumulation in &amp;lt;i&amp;gt;saccharomyces cerevisiae&amp;lt;/i&amp;gt;: comparative study by extraction and DAPI staining

Ryazanova, L. P.; Андреева, Н.А.; Kulakovskaya, T. V.; Valiakhmetov, Airat; Yashin, V. A.; Vagabov, V. M.; Kulaev, I. S.

doi:10.4236/abb.2011.24042

Cited by 7 publications

(4 citation statements)

References 21 publications

(36 reference statements)

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“…DAPI is used at concentrations of 0.1–1 μg DAPI mL –1 for DNA staining and at 3–50 μg DAPI mL –1 for polyP staining. ,,,,− If the cells are treated gently and the microscopic analysis is done quickly after the DAPI staining, only the cell wall-associated polyP can be detected . The polyP concentration can even be quantified within the cell with quantitative image analysis .…”

Section: Microscopymentioning

confidence: 99%

Methods for the Analysis of Polyphosphate in the Life Sciences

2020

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Inorganic polyphosphate (polyP) is the polymer of orthophosphate and can be found in all living organisms. For polyP characterization, one or more of six parameters are of interest: the molecular structure (linear, cyclic, or branched), the concentration, the average chain length, the chain length distribution, the cellular localization, and the cation composition. Here, the merits, limitations, and critical parameters of the state-of-the-art methods for the analysis of the six parameters from the life sciences are discussed. With this contribution, we aim to lower the entry barrier into the analytics of polyP, a molecule with prominent, yet often incompletely understood, contributions to cellular function. See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles. b Refers to absorption and fluorescence spectroscopy. Analytical Chemistry pubs.acs.org/ac Feature https://dx.

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Section: Microscopymentioning

confidence: 99%

Methods for the Analysis of Polyphosphate in the Life Sciences

2020

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“…The cells of both strains of S. cerevisiae were weakly stained by DAPI in the stationary phase independent of the presence or absence of Mn 2+ (Figs 4 and 5b and f). It is noteworthy that the cell of S. cerevisiae does not always show intense fluorescence with DAPI even at high concentrations of PolyP (Ryazanova et al, 2011). The absence of DAPI-fluorescence is not always indicative of the low PolyP level in the yeast, and therefore, chemical extraction of this polymer is still necessary for its quantification.…”

Section: Changes In Cell Morphology and Dapi Staining With Excess Mn 2+mentioning

confidence: 99%

Adaptation ofSaccharomyces cerevisiaeto toxic manganese concentration triggers changes in inorganic polyphosphates

et al. 2013

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The ability of Saccharomyces cerevisiae to adapt to toxic Mn(2+) concentration (4 mM) after an unusually long lag phase has been demonstrated for the first time. The mutants lacking exopolyphosphatase PPX1 did not change the adaptation time, whereas the mutants lacking exopolyphosphatase PPN1 reduced the lag period compared with the wild-type strains. The cell populations of WT and ΔPPN1 in the stationary phase at cultivation with Mn(2+) contained a substantial number of enlarged cells with a giant vacuole. The adaptation correlated with the triggering of polyphosphate metabolism: the drastic increase in the rate and chain length of acid-soluble polyphosphate. The share of this fraction, which is believed to be localized in the cytoplasm, increased to 76%. Its average chain length increased to 200 phosphate residues compared with 15 at the cultivation in the absence of manganese. DAPI-stained inclusions in the cytoplasm were accumulated in the lag phase during the cultivation with Mn(2+).

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“…Indeed, disturbing phosphate homeostasis leads to marked increases in calcium in Saccharomyces [38]. Future work should include analysis by additional assays [39] [40] and in many more vps strains, as well as additional pharmacological agents, such as wortmannin, an inhibitor of phosphoinositide 3-kinases.…”

Section: Pharmacological Agent Analysismentioning

confidence: 99%

Analysis of Volutin Granule Formation in <i>Saccharomyces cerevisiae</i>

Marshall¹,

Rosa²,

Sanchez³

et al. 2014

AiM

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The budding yeast Saccharomyces cerevisiae serves as an effective model organism for many cellular pathways including phosphate transport, accumulation, and storage. In S. cerevisiae, phosphate is actively transported across the plasma membrane via several phosphate carriers and is then transported into the acidic vacuole (roughly equivalent to the mammalian lysosome with degradative functions but with additional storage functions, such as calcium) where it is synthesized into volutin, a storage form of polyphosphate, found in many organisms. We have been studying volutin granule formation in wild type cells to determine the physiological requirements for formation and in mutants to determine the pathway by which the volutin biosynthetic proteins are transported to the vacuole. Undertaking an analysis of volutin formation in yeast vacuoles by blocking vacuole function with pharmacological agents, such as ionomycin and CCCP, we see that vacuole pH as well as vacuolar calcium seems critical for volutin formation. Different blocks in vacuolar protein sorting have differential effects on volutin granule accumulation, with volutin granule formation seen in all mutant strains thus far tested, except for vps33, a mutant cell strain lacking all vacuolar structure. Our data are consistent with pleiotrophic effects of vacuolar physiological function blocks leading to a decrease in volutin formation.

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The early stage of polyphosphate accumulation in <i>saccharomyces cerevisiae</i>: comparative study by extraction and DAPI staining

Abstract: ABSTRACT

Cited by 7 publications

References 21 publications

Methods for the Analysis of Polyphosphate in the Life Sciences

Methods for the Analysis of Polyphosphate in the Life Sciences

Adaptation ofSaccharomyces cerevisiaeto toxic manganese concentration triggers changes in inorganic polyphosphates

Analysis of Volutin Granule Formation in <i>Saccharomyces cerevisiae</i>

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The early stage of polyphosphate accumulation in &lt;i&gt;saccharomyces cerevisiae&lt;/i&gt;: comparative study by extraction and DAPI staining

Abstract: ABSTRACT

Cited by 7 publications

References 21 publications

Methods for the Analysis of Polyphosphate in the Life Sciences

Methods for the Analysis of Polyphosphate in the Life Sciences

Adaptation ofSaccharomyces cerevisiaeto toxic manganese concentration triggers changes in inorganic polyphosphates

Analysis of Volutin Granule Formation in &lt;i&gt;Saccharomyces cerevisiae&lt;/i&gt;

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The early stage of polyphosphate accumulation in <i>saccharomyces cerevisiae</i>: comparative study by extraction and DAPI staining

Analysis of Volutin Granule Formation in <i>Saccharomyces cerevisiae</i>