Ca(2+)-dependent glycolysis activation mediates apoptotic ATP elevation in HeLa cells.
It was previously shown that cells die with increased cytosolic ATP after stimulation with apoptotic inducers including staurosporine (STS). To identify the source of apoptotic ATP elevation, we monitored, in real time, the cytosolic ATP level in luciferase-expressing HeLa cells. A mitochondrial uncoupler or a respiration chain inhibitor was found to decrease cytosolic ATP by about 50%. However, even when mitochondrial ATP synthesis was suppressed, STS induced a profound elevation of intracellular ATP. In contrast, the STS-induced ATP increase was prevented by any of three inhibitors of the glycolytic pathway: 2-deoxyglucose, iodoacetamide, and NaF. The STS effect strongly depended on intracellular calcium and was mimicked by a calcium ionophore. We conclude that Ca(2+)-dependent activation of anaerobic glycolysis, but not aerobic mitochondrial oxidative phosphorylation, is responsible for the STS-induced elevation of ATP in apoptotic HeLa cells.
Zamaraeva
Maria V
MV
Department of Cell Physiology, National Institute for Physiological Sciences, Okazaki 444-8585, Japan.
Sabirov
Ravshan Z
RZ
Manabe
Ken-ichi
K
Okada
Yasunobu
Y
D000255:Adenosine Triphosphate / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D003847:Deoxyglucose / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007460:Iodoacetamide / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D010085:Oxidative Phosphorylation / Q000187:drug effects; D010416:Pentachlorophenol / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D014162:Transfection; D014475:Uncoupling Agents / Q000494:pharmacology
D002614:Chelating Agents; D007476:Ionophores; D014475:Uncoupling Agents; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D015759:Ionomycin; D000255:Adenosine Triphosphate; D012969:Sodium Fluoride; D003847:Deoxyglucose; D010416:Pentachlorophenol; D008156:Luciferases; D019311:Staurosporine; D002118:Calcium; D007460:Iodoacetamide