Regulation of N-linked glycoprotein assembly in uteri by steroid hormones.
The effects of the steroid hormones 17 beta-estradiol (E2) and progesterone on N-linked glycoprotein assembly in ovariectomized mice have been examined. Both priming and nidatory E2 markedly stimulate [3H]mannose incorporation (3- to 6-fold) into uterine glycoproteins, whereas uterine bulk protein synthesis is not stimulated under the same conditions. Progesterone alone stimulates glycoprotein synthesis modestly (1.5-fold) over that in oil-injected controls, but antagonizes the action of E2 when coinjected with the estrogen. The E2 effect is not systemic, because livers from these same animals do not display an increase in glycoprotein synthesis. When mice were injected with tamoxifen or clomiphene, two drugs that mimic E2 actions in uteri without inducing the full extent of cell proliferation that normally accompanies E2 treatment, a similar enhancement of uterine glycoprotein synthesis was observed. Although mannosylphosphoryldolichol synthase activity rose in parallel with glycoprotein synthesis during E2 priming, the apparent activities of two other enzymes involved in the assembly of N-linked glycoproteins, namely chitobiosylpyrophosphoryldolichol synthase and oligosaccharyltransferase, remained relatively unchanged. Furthermore, the apparent in vivo rate of dolichol phosphorylation was not altered during E2 priming. Supplementation of uterine tissue slices with dolichylphosphate failed to enhance the rate of protein glycosylation in vivo. In addition, changes in the pool sizes of GDP-mannose did not correlate with changes in the in vivo rate of glycoprotein synthesis. Collectively, these observations indicate that the E2-dependent increase in glycoprotein synthesis is not likely to be due to increased enzyme activities for oligosaccharide assembly or transfer to protein, increased dolichylphosphate availability, or increased sugar nucleotide availability. To study the effects of E2 on the production of specific glycoproteins, the pattern of [3H]mannose-labeled glycoproteins produced as a function of days of E2 priming was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Estrogen priming induced the secretion of 9-11 [3H]mannose-labeled glycoproteins by uteri; however, the pattern of tissue-associated glycoproteins remained constant throughout this interval. It appears, therefore, that estrogen priming induces the secretion of a few specific glycoproteins while generally enhancing the production of most tissue-associated glycoproteins. Most (70%) of the [3H]mannose-labeled oligosaccharide chains of these glycoproteins were of the polymannose type.(ABSTRACT TRUNCATED AT 400 WORDS)
A
A
Dutt
J P
JP
Tang
J K
JK
Welply
D D
DD
Carson
D000818:Animals; D002236:Carbohydrate Conformation; D002850:Chromatography, Gel; D002996:Clomiphene / Q000494:pharmacology; D004286:Dolichols / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010064:Embryo Implantation; D004958:Estradiol / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D005964:Glucosyltransferases / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis*; D006155:Guanosine Diphosphate Mannose / Q000378:metabolism; D006602:Hexosyltransferases; D007700:Kinetics; D008358:Mannose / Q000378:metabolism; D008364:Mannosyltransferases / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D008970:Molecular Weight; D017351:N-Acetylglucosaminyltransferases; D009844:Oligosaccharides / Q000378:metabolism; D010052:Ovariectomy; D010766:Phosphorylation; D011374:Progesterone / Q000008:administration & dosage / Q000494:pharmacology*; D013629:Tamoxifen / Q000494:pharmacology; D014166:Transferases / Q000378:metabolism; D014599:Uterus / Q000187:drug effects / Q000378:metabolism*
D004286:Dolichols; D006023:Glycoproteins; D008565:Membrane Proteins; D009844:Oligosaccharides; D013629:Tamoxifen; D002996:Clomiphene; D006155:Guanosine Diphosphate Mannose; D011374:Progesterone; D004958:Estradiol; D014166:Transferases; D005964:Glucosyltransferases; D006602:Hexosyltransferases; D008364:Mannosyltransferases; D017351:N-Acetylglucosaminyltransferases; C047836:chitobiosylpyrophosphoryldolichol synthase; C047834:N-acetylglucosaminyldiphosphodolichol N-acetylglucosaminyltransferase; C047835:dolichyl-phosphate alpha-N-acetylglucosaminyltransferase; C022755:dolichyl-phosphate beta-D-mannosyltransferase; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase; D008358:Mannose