Changes in composition of newly synthesized sphingolipids of HeLa cells during the cell cycle -- suppression of sphingomyelin and higher-glycosphingolipid synthesis and accumulation of ceramide and glucosylceramide in mitotic cells.
Sphingolipid biosynthesis in synchronized HeLa cells was studied by pulse labeling with [14C]Ser or [14C]Gal and a simple TLC method. The major HeLa cell sphingolipids are ceramide (Cer), sphingomyelin, glucosylceramide (GlcCer), lactosylceramide (LacCer), globotriaosylceramide (Gb3Cer), N-acetylneuraminosylgangl iotriaosylceramide (GM2) and sialylparagloboside (G[M1-GlcNAc]). The sphingolipid biosynthetic profiles of HeLa cells in the G1, G1/S boundary, S and G2 phases were similar, but significant changes occurred during M phase, when incorporation of radioactivity into sphingomyelin, Gb3Cer and a mixture of GM2 and G(M1-GlcNAc) decreased, and those of Cer and GlcCer increased. These data indicate that transfer of phosphocholine and galactose to Cer and GlcCer, respectively, decreased in mitotic cells, resulting in accumulation of Cer and GlcCer. Analysis of LacCer synthase activity revealed that GlcCer accumulation was not due to reduced activity of this enzyme. The results suggest that Cer and GlcCer accumulation in mitotic cells resulted from suppression of sphingomyelin and LacCer synthesis, probably caused by vesiculation of membranous organelles, such as the endoplasmic reticulum and Golgi apparatus.
Yokoyama
K
K
Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
Suzuki
M
M
Kawashima
I
I
Karasawa
K
K
Nojima
S
S
Enomoto
T
T
Tai
T
T
Suzuki
A
A
Setaka
M
M
D001345:Autoradiography; D002250:Carbon Radioisotopes; D002453:Cell Cycle / Q000502:physiology*; D002518:Ceramides / Q000096:biosynthesis; D002855:Chromatography, Thin Layer; D005700:Galactosyltransferases / Q000378:metabolism; D005963:Glucosylceramides / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000378:metabolism; D006028:Glycosphingolipids / Q000096:biosynthesis; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008938:Mitosis; D013107:Sphingolipids / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013109:Sphingomyelins / Q000096:biosynthesis; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism
D002250:Carbon Radioisotopes; D002518:Ceramides; D005963:Glucosylceramides; D006028:Glycosphingolipids; D013107:Sphingolipids; D013109:Sphingomyelins; D005700:Galactosyltransferases; D005964:Glucosyltransferases; C039090:UDPgalactose-glucosylceramide galactosyltransferase; C023246:ceramide glucosyltransferase; D017855:Transferases (Other Substituted Phosphate Groups); C019863:ceramide cholinephosphotransferase