Ceramide synthases 2, 5, and 6 confer distinct roles in radiation-induced apoptosis in HeLa cells.
The role of ceramide neo-genesis in cellular stress response signaling is gaining increasing attention with recent progress in elucidating the novel roles and biochemical properties of the ceramide synthase (CerS) enzymes. Selective tissue and subcellular distribution of the six mammalian CerS isoforms, combined with distinct fatty acyl chain length substrate preferences, implicate differential functions of specific ceramide species in cellular signaling. We report here that ionizing radiation (IR) induces de novo synthesis of ceramide to influence HeLa cell apoptosis by specifically activating CerS isoforms 2, 5, and 6 that generate opposing anti- and pro-apoptotic ceramides in mitochondrial membranes. Overexpression of CerS2 resulted in partial protection from IR-induced apoptosis whereas overexpression of CerS5 increased apoptosis in HeLa cells. Knockdown studies determined that CerS2 is responsible for all observable IR-induced C(24:0) CerS activity, and while CerS5 and CerS6 each confer approximately 50% of the C(16:0) CerS baseline synthetic activity, both are required for IR-induced activity. Additionally, co-immunoprecipitation studies suggest that CerS2, 5, and 6 might exist as heterocomplexes in HeLa cells, providing further insight into the regulation of CerS proteins. These data add to the growing body of evidence demonstrating interplay among the CerS proteins in a stress stimulus-, cell type- and subcellular compartment-specific manner.
Mesicek
Judith
J
Laboratory of Signal Transduction, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
Lee
Hyunmi
H
Feldman
Taya
T
Jiang
Xuejun
X
Skobeleva
Anastasia
A
Berdyshev
Evgeny V
EV
Haimovitz-Friedman
Adriana
A
Fuks
Zvi
Z
Kolesnick
Richard
R
D000214:Acyl Coenzyme A / Q000378:metabolism; D017209:Apoptosis; D002518:Ceramides / Q000096:biosynthesis; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism* / Q000502:physiology; D008928:Mitochondria / Q000378:metabolism / Q000528:radiation effects; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D011839:Radiation, Ionizing; D051049:Sphingosine N-Acyltransferase / Q000378:metabolism* / Q000502:physiology; D025521:Tumor Suppressor Proteins / Q000378:metabolism* / Q000502:physiology
D000214:Acyl Coenzyme A; D002518:Ceramides; D007527:Isoenzymes; D008565:Membrane Proteins; D025521:Tumor Suppressor Proteins; D010171:Palmitoyl Coenzyme A; C018582:lignoceroyl-coenzyme A; C438311:CERS2 protein, human; C505722:CERS5 protein, human; C506123:CERS6 protein, human; D051049:Sphingosine N-Acyltransferase
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