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Influence of S-adenosylmethionine on DAPI-induced fluorescence of polyphosphate in the yeast vacuole
Abstract:
Use of the fluorochrome 4',6-diamidino-2-phenylindole.2 HCl (DAPI) in ultraviolet microscopy revealed fluorescent objects in Brownian motion within the vacuoles of seven species of yeast. The abundance of these bodies increased when cells of Saccharomyces cerevisiae were transferred from growth medium to a glucose-phosphate solution, indicating that they contain polyphosphate. In addition, the effect on vacuolar fluorescence of supplementing a defined growth medium with amino acids provided evidence that they also contained S-adenosylmethionine. These deductions were supported by in vitro studies of the interaction and fluorescence of polyphosphate, S-adenosylmethionine, and DAPI. Vacuolar fluorescence of cells in suspension in flucose-phosphate solution was less after addition of exogenous arginine, lysine, or glutamine but not after addition of alanine, aspartic acid, or methionine. Mithramycin was superior to DAPI as a fluorochrome for ultraviolet demonstration of yeast nuclei since it stained the nuclei much more intensely and did not fluoresce with other material in the cells.
Insights
This study reveals fluorescent bodies in yeast vacuoles containing polyphosphate and S-adenosylmethionine. Their abundance changes with nutrient conditions, offering insights into yeast cell metabolism.
Area of Science:
- Microbiology
- Cell Biology
- Biochemistry
Background:
- Vacuoles in yeast cells are dynamic organelles.
- Fluorescent probes are crucial for visualizing intracellular components.
Purpose of the Study:
- To identify the composition of fluorescent bodies within yeast vacuoles.
- To investigate the role of polyphosphate and S-adenosylmethionine in yeast vacuoles.
- To compare the efficacy of DAPI and mithramycin as fluorochromes for yeast cell imaging.
Main Methods:
- Ultraviolet microscopy using 4',6-diamidino-2-phenylindole.2 HCl (DAPI) fluorochrome.
- Culturing Saccharomyces cerevisiae in different media (growth, glucose-phosphate).
- Supplementation of growth media with specific amino acids.
- In vitro studies of fluorochrome interactions with polyphosphate and S-adenosylmethionine.
Main Results:
- DAPI revealed fluorescent bodies in yeast vacuoles exhibiting Brownian motion.
- Abundance of these bodies increased in glucose-phosphate solution, suggesting polyphosphate content.
- Supplementation with certain amino acids altered vacuolar fluorescence, indicating S-adenosylmethionine presence.
- Mithramycin proved more effective than DAPI for yeast nuclear staining.
Conclusions:
- Yeast vacuoles contain polyphosphate and S-adenosylmethionine, with their levels influenced by nutrient availability.
- DAPI is useful for visualizing these vacuolar components, while mithramycin is superior for nuclear staining.