Genomic Analysis of ATP Efflux in Saccharomyces cerevisiae

Abstract

Adenosine triphosphate (ATP) plays an important role as a primary molecule for the transfer of chemical energy to drive biological processes. ATP also functions as an extracellular signaling molecule in a diverse array of eukaryotic taxa in a conserved process known as purinergic signaling. Given the important roles of extracellular ATP in cell signaling, we sought to comprehensively elucidate the pathways and mechanisms governing ATP efflux from eukaryotic cells. Here, we present results of a genomic analysis of ATP efflux from Saccharomyces cerevisiae by measuring extracellular ATP levels in cultures of 4609 deletion mutants. This screen revealed key cellular processes that regulate extracellular ATP levels, including mitochondrial translation and vesicle sorting in the late endosome, indicating that ATP production and transport through vesicles are required for efflux. We also observed evidence for altered ATP efflux in strains deleted for genes involved in amino acid signaling, and mitochondrial retrograde signaling. Based on these results, we propose a model in which the retrograde signaling pathway potentiates amino acid signaling to promote mitochondrial respiration. This study advances our understanding of the mechanism of ATP secretion in eukaryotes and implicates TOR complex 1 (TORC1) and nutrient signaling pathways in the regulation of ATP efflux. These results will facilitate analysis of ATP efflux mechanisms in higher eukaryotes.

ATP Mediates Yeast Cell-Cell Communication

“…Although yeast in synthetic media generate 1-2 uM ATP after 40 hours, in a path regulated by the EGO complex (Peters et al, 2016), the lesser amount under our experimental conditions (Fig. 10) appears to be discrepant with finding that ≥1 uM ATP concentrations was required to generate a detectable stimulation of growth on agar (Figs.…”

Draught beer hygiene: cleaning of dispense tap nozzles

“…and accuracy of plate counts (selective media, non‐culturability, homogeneity of sampling, etc.) To add further complexity, there are also reports of ATP efflux from yeast where it has been implicated in signalling and bacteria where it is linked to active growth . Nevertheless, ATP bioluminescence has transformed hygiene quality assurance and is integral to real‐time monitoring of cleaning in place (CiP) via testing of surfaces or rinses.…”

Extracellular ATP released fromCandida albicansactivates non-peptidergic neurons to augment host defense

“…Furthermore, eATP levels were independent of levels of intracellular ATP, which is well known to be essential for growth, filament formation, and successful invasion [22]. ATP is released by S. cerevisiae and a mutational analysis revealed that many genes related to the secretory pathways were required for ATP efflux [27]. This suggests that release of eATP occurs through a defined mechanism and is not unique to C. albicans.…”

“…We focused exclusively on mutants with reduced eATP since the predicted increased levels of infection would be unlikely to result from the effects of genetic mutation that affect other aspects of C. albicans biology. A library screen of S. cerevisiae identified several genes required for release of eATP [27]. The majority of available C. albicans orthologous mutant strains, however, were on genetic background that did not release eATP (e.g.…”

“…As predicted, on day +3 following epicutaneous infection, CFUs of the hda1 Δ/Δ mutant were significantly higher than those from control J4-2.1 cells ( Fig 2H). Finally, we screened a series of dox-inducible OE strains based on our prediction from the data in Peters et al (2016) that they would be affected in eATP secretion [28,29]We found two clones that secreted appreciable amounts of eATP and showed similar growth kinetics. Incubation with 50 μg/ml of doxycycline reduced eATP release from P TET -ORF19.2898 but not from P TET -O-ORF19.185 (Fig 2I-J).…”