Sphingosine-phosphate lyase enhances stress-induced ceramide generation and apoptosis.
Sphingosine-1-phosphate lyase is a widely expressed enzyme that catalyzes the essentially irreversible cleavage of the signaling molecule sphingosine 1-phosphate. To investigate whether sphingosine-1-phosphate lyase influences mammalian cell fate decisions, a recombinant human sphingosine-1-phosphate lyase fused to green fluorescent protein was expressed in HEK293 cells. The recombinant enzyme was active, localized to the endoplasmic reticulum, and reduced baseline sphingosine and sphingosine 1-phosphate levels. Stable overexpression led to diminished viability under stress, which was attributed to an increase in apoptosis and was reversible in a dose-dependent manner by exogenous sphingosine 1-phosphate. In contrast to sphingosine 1-phosphate, the products of the lyase reaction had no effect on apoptosis. Lyase enzymatic activity was required to potentiate apoptosis, because cells expressing a catalytically inactive enzyme behaved like controls. Stress increased the amounts of long- and very long-chain ceramides in HEK293 cells, and this was enhanced in cells overexpressing wild type but not catalytically inactive lyase. The ceramide increases appeared to be required for apoptosis, because inhibition of ceramide synthase with fumonisin B1 decreased apoptosis in lyase-overexpressing cells. Thus, sphingosine-1-phosphate lyase overexpression in HEK293 cells decreases sphingosine and sphingosine 1-phosphate amounts but elevates stress-induced ceramide generation and apoptosis. This identifies sphingosine-1-phosphate lyase as a dual modulator of sphingosine 1-phosphate and ceramide metabolism as well as a regulator of cell fate decisions and, hence, a potential target for diseases with an imbalance in these biomodulators, such as cancer.
Children's Hospital Oakland Research Institute, Oakland, California 94609-1673, USA.
Ulrike
U
Reiss
Babak
B
Oskouian
Jianhui
J
Zhou
Vinita
V
Gupta
Prathap
P
Sooriyakumaran
Samuel
S
Kelly
Elaine
E
Wang
Alfred H
AH
Merrill
Julie D
JD
Saba
D000446:Aldehyde-Lyases / Q000737:chemistry*; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002460:Cell Line; D002518:Ceramides / Q000378:metabolism*; D003001:Cloning, Molecular; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009154:Mutation; D011994:Recombinant Proteins / Q000737:chemistry; D013107:Sphingolipids / Q000378:metabolism; D013110:Sphingosine / Q000378:metabolism; D013312:Stress, Physiological; D013997:Time Factors; D014162:Transfection
D002518:Ceramides; D018076:DNA, Complementary; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D013107:Sphingolipids; D049452:Green Fluorescent Proteins; D045304:Cytochromes c; D004247:DNA; D000446:Aldehyde-Lyases; C022649:sphingosine 1-phosphate lyase (aldolase); D013110:Sphingosine