Externalization of phosphatidylserine during apoptosis does not specifically require either isoform of phosphatidylserine synthase.
Phosphatidylserine (PtdSer) is made in mammalian cells by two PtdSer synthases, PSS1 and PSS2. In the plasma membrane PtdSer is normally localized on the inner leaflet but undergoes transbilayer movement during apoptosis and becomes exposed on the cell surface. We induced apoptosis with staurosporine in four Chinese hamster ovary (CHO) cell lines that are deficient in PSS1 and/or PSS2 to determine if PtdSer generated by either of these enzymes is required for externalization on the cell surface during apoptosis. The onset of apoptosis was confirmed by the appearance of morphological changes and DNA fragmentation while the plasma membrane remained largely intact. In all cell lines, regardless of their content of PSS1 and/or PSS2, apoptosis occurred to approximately the same extent, and within approximately the same time frame, as in parental CHO-K1 cells. The exposure of PtdSer on the cell surface was assessed by annexin V labeling and flow cytometry. Cells that were deficient in either PSS1 or PSS2, as well as cells that were deficient in both PSS1 and PSS2, externalized normal amounts of PtdSer. Our study demonstrates, that reduction of in vitro serine-exchange activity, even by 97%, does not restrict the externalization of PtdSer during apoptosis. Moreover, a normal level of expression of PSS1 and/or PSS2 is not required for generating the pool of PtdSer externalized during apoptosis.
CIHR Group on the Molecular and Cell Biology of Lipids and Department of Medicine, 332 HMRC, University of Alberta, Edmonton, AB, Canada T6G 2S2.
Paul A
PA
Grandmaison
Terry S
TS
Nanowski
Jean E
JE
Vance
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000172:deficiency / Q000378:metabolism*; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D007527:Isoenzymes / Q000172:deficiency / Q000378:metabolism; D045022:Lactate Dehydrogenases / Q000032:analysis; D010718:Phosphatidylserines / Q000032:analysis / Q000378:metabolism*; D019311:Staurosporine
D007527:Isoenzymes; D010718:Phosphatidylserines; D045022:Lactate Dehydrogenases; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase; D019311:Staurosporine