Cell surface and Golgi pools of beta-1,4-galactosyltransferase are differentially regulated during embryonal carcinoma cell differentiation.
beta-1,4-Galactosyltransferase (GalTase) has two functionally distinct subcellular distributions. In the Golgi apparatus, GalTase participates in the glycosylation of secretory and membrane-bound glycoproteins, whereas on the cell surface it mediates specific aspects of intercellular adhesion. For this study, a murine GalTase clone was obtained by screening a lambda gt10 cDNA library made from F9 embryonal carcinoma cells with a heterologous bovine GalTase cDNA probe. The murine GalTase cDNA probe was used in conjunction with assays of GalTase activity to investigate the expression and distribution of GalTase during differentiation of F9 stem cells into secretory endodermal epithelium. During the initial phase of F9 cell differentiation, GalTase mRNA levels remained relatively constant; however, as differentiation progressed, as assayed by expression of the differentiation-specific marker laminin B1, GalTase mRNA levels and enzyme activity rose dramatically. Furthermore, subcellular fractionation of these cells showed that the increased GalTase levels were specifically associated with the Golgi apparatus, whereas GalTase specific activity on the plasma membrane remained constant. These results show that levels of cell surface and Golgi GalTase change relative to one another during F9 cell differentiation and suggest that these functionally distinct pools of GalTase are independently and differentially regulated.
Lopez
L C
LC
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Maillet
C M
CM
Oleszkowicz
K
K
Shur
B D
BD
D015152:Blotting, Northern; D002454:Cell Differentiation; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D015342:DNA Probes; D054278:Embryonal Carcinoma Stem Cells; D005819:Genetic Markers; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D007788:Lactose Synthase / Q000096:biosynthesis*; D007797:Laminin / Q000096:biosynthesis / Q000235:genetics; D009237:N-Acetyllactosamine Synthase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000201:enzymology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D015183:Restriction Mapping
D015342:DNA Probes; D005819:Genetic Markers; D007797:Laminin; D012333:RNA, Messenger; D007788:Lactose Synthase; D009237:N-Acetyllactosamine Synthase