ArticleProceedings of the National Academy of Sciences of the United States of America1991
Monitoring of intracellular calcium in Saccharomyces cerevisiae with an apoaequorin cDNA expression system.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1991. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed, 124 citations in OpenAlex.
- KCS1 and VIP1, the genes encoding yeast phosphoinositol pyrophosphate synthases, are required for CaFEBS open bio · 2025Article
- Calcium Signaling Is a Universal Carbon Source Signal Transducer and Effects an Ionic Memory of Past Carbon Sources.International journal of molecular sciences · 2025Article
- Shedding Light on Calcium Dynamics in the Budding Yeast: A Review on Calcium Monitoring with Recombinant Aequorin.Molecules (Basel, Switzerland) · 2024Review
- Article
- Overexpression profiling reveals cellular requirements in the context of genetic backgrounds and environments.PLoS genetics · 2023Article
- Rational engineering approaches for establishing insect olfaction reporters in yeast.Biotechnology notes (Amsterdam, Netherlands) · 2023Article
- The GEM-GECO Calcium Indicator Is Useable inInternational journal of molecular sciences · 2022Article
- Calmodulin kinase 2 genetically interacts with Rch1p to negatively regulate calcium import into Saccharomyces cerevisiae after extracellular calcium pulse.Archives of microbiology · 2022Article
- Cytotoxicity of Oleandrin Is Mediated by Calcium Influx and by Increased Manganese Uptake inMolecules (Basel, Switzerland) · 2020Article
- Yeast-model-based study identified myosin- and calcium-dependent calmodulin signalling as a potential target for drug intervention in chorea-acanthocytosis.Disease models & mechanisms · 2019Article
- Article
- Sexual pheromone modulates the frequency of cytosolic CaMolecular biology of the cell · 2017Article
- Glutathione depletion activates the yeast vacuolar transient receptor potential channel, Yvc1p, by reversible glutathionylation of specific cysteines.Molecular biology of the cell · 2016Article
- Calcium signaling and copper toxicity in Saccharomyces cerevisiae cells.Environmental science and pollution research international · 2016Article
- Interaction between lanthanide ions and Saccharomyces cerevisiae cells.Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · 2015Article
- Regulation of cation balance in Saccharomyces cerevisiae.Genetics · 2013Review
- A yeast expression system for functional and pharmacological studies of the malaria parasite Ca²⁺/H⁺ antiporter.Malaria journal · 2012Article
- Involvement of the putative Ca²⁺-permeable mechanosensitive channels, NtMCA1 and NtMCA2, in Ca²⁺ uptake, Ca²⁺-dependent cell proliferation and mechanical stress-induced gene expression in tobacco (Nicotiana tabacum) BY-2 cells.Journal of plant research · 2012Article
- Illuminating cellular physiology: recent developments.Science progress · 2007Review
- In Saccharomyces cerevisiae, cations control the fate of the energy derived from oxidative metabolism through the opening and closing of the yeast mitochondrial unselective channel.Journal of bioenergetics and biomembranes · 2003Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A method is described for measuring cytosolic free Ca2+ and its time-dependent changes in the yeast Saccharomyces cerevisiae by using the luminescent protein aequorin as a Ca(2+)-specific indicator. This method with intact yeast cells is labeled "in vivo" to distinguish it from methods with cell extracts, labeled "in vitro." A plasmid in which the apoaequorin cDNA was joined downstream from the glyceraldehyde-3-phosphate dehydrogenase gene promoter was constructed and introduced into yeast cells. The intracellular concentration of apoaequorin expressed by the cDNA was approximately 1 microM, which was high enough to detect the cytosolic Ca2+. Growth of the transformed cells was normal. In the in vitro method, apoaequorin in crude cell extracts was regenerated into aequorin by mixing with coelenterazine, the substrate for the luminescence reaction, whereas in the in vivo method, aequorin was regenerated by incubating intact cells with coelenterazine. Simultaneous addition of 10 mM CaCl2 and 10 microM A23187, a Ca2+ ionophore, to coelenterazine-incorporated cells generated luminescence. Coelenterazine-incorporated cells also responded to native extracellular stimuli. A mating pheromone, alpha-factor, added to cells of mating type a or alpha, generated extracellular Ca(2+)-dependent luminescence specifically in a mating type cells, with maximal intensity occurring 45-50 min after addition of alpha-factor. Glucose added to glucose-starved G0/G1 cells stimulated an increase in extracellular Ca(2+)-dependent luminescence with maximal intensity occurring 2 min after addition. These results show the usefulness of the aequorin system in monitoring [Ca2+]i response to extracellular stimuli in yeast cells.
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