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. 1996 Jun 1;316(Pt 2):575–581. doi: 10.1042/bj3160575

Long-term effect of cyclic AMP on N-glycosylation is caused by an increase in the activity of the cis-prenyltransferase.

M Konrad 1, W E Merz 1
PMCID: PMC1217387  PMID: 8687403

Abstract

Previously we have shown that long-term pretreatment of JEG-3 choriocarcinoma cells with 8-bromo-cAMP increases the capacity for N-glycosylation that was caused by an 8-10-fold enlargement of the dolichol pyrophosphoryl oligosaccharide (Dol-PP-oligosaccharide) pool [Konrad and Merz (1994) J. Biol. Chem. 269, 8659-8666]. The factors involved in the effect of cAMP on synthesis of Dol-PP-oligosaccharide are investigated here. The GlcNAc transfer to dolichol phosphate (Dol-P) was found to be unaffected by pretreatment with 8-bromo-cAMP. By measuring the uptake of [3H]mevalonate, a 20-fold increase in the incorporation of the label into Dol-P was observed in the cells treated with 8-bromo-cAMP. Under the same conditions, the synthesis of dolichol was enhanced 60-fold. However, the incorporation of the radioactivity into cholesterol was not increased in the JEG-3 cells pretreated with 8-bromo-cAMP, which suggests a specific stimulation of the dolichol/Dol-P pathway by cAMP. The cis-prenyltransferase activity was found to be increased 10-fold in cells pretreated with 8-bromo-cAMP. Dolichol kinase activity was unaffected by stimulation with 8-bromo-cAMP. The present study suggests that the larger glycosylation capacity in JEG-3 cells treated with 8-bromo-cAMP is caused by an increase in the microsomal cis-prenyltransferase activity.

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Selected References

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  1. Abeijon C., Hirschberg C. B. Topography of glycosylation reactions in the endoplasmic reticulum. Trends Biochem Sci. 1992 Jan;17(1):32–36. doi: 10.1016/0968-0004(92)90424-8. [DOI] [PubMed] [Google Scholar]
  2. Adair W. L., Jr, Cafmeyer N. Characterization of the Saccharomyces cerevisiae cis-prenyltransferase required for dolichyl phosphate biosynthesis. Arch Biochem Biophys. 1987 Dec;259(2):589–596. doi: 10.1016/0003-9861(87)90525-x. [DOI] [PubMed] [Google Scholar]
  3. Adair W. L., Jr, Cafmeyer N., Keller R. K. Solubilization and characterization of the long chain prenyltransferase involved in dolichyl phosphate biosynthesis. J Biol Chem. 1984 Apr 10;259(7):4441–4446. [PubMed] [Google Scholar]
  4. Adair W. L., Keller R. K. Isolation and assay of dolichol and dolichyl phosphate. Methods Enzymol. 1985;111:201–215. doi: 10.1016/s0076-6879(85)11010-4. [DOI] [PubMed] [Google Scholar]
  5. Appelkvist E. L., Kalén A. Biosynthesis of dolichol by rat liver peroxisomes. Eur J Biochem. 1989 Nov 20;185(3):503–509. doi: 10.1111/j.1432-1033.1989.tb15142.x. [DOI] [PubMed] [Google Scholar]
  6. Astrand I. M., Fries E., Chojnacki T., Dallner G. Inhibition of dolichyl phosphate biosynthesis by compactin in cultured rat hepatocytes. Eur J Biochem. 1986 Mar 3;155(2):447–452. doi: 10.1111/j.1432-1033.1986.tb09511.x. [DOI] [PubMed] [Google Scholar]
  7. Baba T., Morris C., Allen C. M. Dehydrodolichyl diphosphate synthetase from rat seminiferous tubules. Arch Biochem Biophys. 1987 Feb 1;252(2):440–450. doi: 10.1016/0003-9861(87)90050-6. [DOI] [PubMed] [Google Scholar]
  8. Banerjee D. K., Kousvelari E. E., Baum B. J. beta-Adrenergic activation of glycosyltransferases in the dolichylmonophosphate-linked pathway of protein N-glycosylation. Biochem Biophys Res Commun. 1985 Jan 16;126(1):123–129. doi: 10.1016/0006-291x(85)90580-7. [DOI] [PubMed] [Google Scholar]
  9. Banerjee D. K., Kousvelari E. E., Baum B. J. cAMP-mediated protein phosphorylation of microsomal membranes increases mannosylphosphodolichol synthase activity. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6389–6393. doi: 10.1073/pnas.84.18.6389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Baudhuin P., Beaufay H., Rahman-Li Y., Sellinger O. Z., Wattiaux R., Jacques P., De Duve C. Tissue fractionation studies. 17. Intracellular distribution of monoamine oxidase, aspartate aminotransferase, alanine aminotransferase, D-amino acid oxidase and catalase in rat-liver tissue. Biochem J. 1964 Jul;92(1):179–184. doi: 10.1042/bj0920179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Bauer P. I., Várady G. A simple method for synthesizing [gamma-32P]nucleoside triphosphates using [32P]acetylphosphate and acetate kinase. Anal Biochem. 1978 Dec;91(2):613–617. doi: 10.1016/0003-2697(78)90547-x. [DOI] [PubMed] [Google Scholar]
  12. Beaufay H., Amar-Costesec A., Feytmans E., Thinès-Sempoux D., Wibo M., Robbi M., Berthet J. Analytical study of microsomes and isolated subcellular membranes from rat liver. I. Biochemical methods. J Cell Biol. 1974 Apr;61(1):188–200. doi: 10.1083/jcb.61.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Burton W. A., Lucas J. J., Waechter C. J. Enhanced chick oviduct dolichol kinase activity during estrogen-induced differentiation. J Biol Chem. 1981 Jan 25;256(2):632–635. [PubMed] [Google Scholar]
  14. Carson D. D., Earles B. J., Lennarz W. J. Enhancement of protein glycosylation in tissue slices by dolichylphosphate. J Biol Chem. 1981 Nov 25;256(22):11552–11557. [PubMed] [Google Scholar]
  15. Carson D. D., Farrar J. D., Laidlaw J., Wright D. A. Selective activation of the N-glycosylation apparatus in uteri by estrogen. J Biol Chem. 1990 Feb 15;265(5):2947–2955. [PubMed] [Google Scholar]
  16. Carson D. D., Tang J. P., Hu G. Estrogen influences dolichyl phosphate distribution among glycolipid pools in mouse uteri. Biochemistry. 1987 Mar 24;26(6):1598–1606. doi: 10.1021/bi00380a017. [DOI] [PubMed] [Google Scholar]
  17. Crick D. C., Scocca J. R., Rush J. S., Frank D. W., Krag S. S., Waechter C. J. Induction of dolichyl-saccharide intermediate biosynthesis corresponds to increased long chain cis-isoprenyltransferase activity during the mitogenic response in mouse B cells. J Biol Chem. 1994 Apr 8;269(14):10559–10565. [PubMed] [Google Scholar]
  18. Crick D. C., Waechter C. J. Long-chain cis-isoprenyltransferase activity is induced early in the developmental program for protein N-glycosylation in embryonic rat brain cells. J Neurochem. 1994 Jan;62(1):247–256. doi: 10.1046/j.1471-4159.1994.62010247.x. [DOI] [PubMed] [Google Scholar]
  19. Dutt A., Tang J. P., Welply J. K., Carson D. D. Regulation of N-linked glycoprotein assembly in uteri by steroid hormones. Endocrinology. 1986 Feb;118(2):661–673. doi: 10.1210/endo-118-2-661. [DOI] [PubMed] [Google Scholar]
  20. Ericsson J., Appelkvist E. L., Thelin A., Chojnacki T., Dallner G. Isoprenoid biosynthesis in rat liver peroxisomes. Characterization of cis-prenyltransferase and squalene synthetase. J Biol Chem. 1992 Sep 15;267(26):18708–18714. [PubMed] [Google Scholar]
  21. Ericsson J., Thelin A., Chojnacki T., Dallner G. Substrate specificity of cis-prenyltransferase in rat liver microsomes. J Biol Chem. 1992 Sep 25;267(27):19730–19735. [PubMed] [Google Scholar]
  22. Friedman G., Chajek-Shaul T., Stein O., Noe L., Etienne J., Stein Y. Beta-adrenergic stimulation enhances translocation, processing and synthesis of lipoprotein lipase in rat heart cells. Biochim Biophys Acta. 1986 Jun 11;877(1):112–120. doi: 10.1016/0005-2760(86)90125-6. [DOI] [PubMed] [Google Scholar]
  23. Grant S. R., Lennarz W. J. Relationship between oligosaccharide-lipid synthesis and protein synthesis in mouse LM cells. Eur J Biochem. 1983 Aug 15;134(3):575–583. doi: 10.1111/j.1432-1033.1983.tb07605.x. [DOI] [PubMed] [Google Scholar]
  24. Hartl F. U., Just W. W., Köster A., Schimassek H. Improved isolation and purification of rat liver peroxisomes by combined rate zonal and equilibrium density centrifugation. Arch Biochem Biophys. 1985 Feb 15;237(1):124–134. doi: 10.1016/0003-9861(85)90261-9. [DOI] [PubMed] [Google Scholar]
  25. Hayes G. R., Lucas J. J. Stimulation of lipid-linked oligosaccharide assembly during oviduct differentiation. J Biol Chem. 1983 Dec 25;258(24):15095–15100. [PubMed] [Google Scholar]
  26. Konrad M., Merz W. E. Regulation of N-glycosylation. Long term effect of cyclic AMP mediates enhanced synthesis of the dolichol pyrophosphate core oligosaccharide. J Biol Chem. 1994 Mar 25;269(12):8659–8666. [PubMed] [Google Scholar]
  27. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  28. Kousvelari E. E., Grant S. R., Baum B. J. beta-Adrenergic receptor regulation of N-linked protein glycosylation in rat parotid acinar cells. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7146–7150. doi: 10.1073/pnas.80.23.7146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  30. Lehrman M. A. Biosynthesis of N-acetylglucosamine-P-P-dolichol, the committed step of asparagine-linked oligosaccharide assembly. Glycobiology. 1991 Dec;1(6):553–562. doi: 10.1093/glycob/1.6.553. [DOI] [PubMed] [Google Scholar]
  31. Lennarz W. J. Glycoprotein synthesis and embryonic development. CRC Crit Rev Biochem. 1983;14(4):257–272. doi: 10.3109/10409238309102795. [DOI] [PubMed] [Google Scholar]
  32. Lucas J. J., Levin E. Increase in the lipid intermediate pathway of protein glycosylation during hen oviduct differentiation. J Biol Chem. 1977 Jun 25;252(12):4330–4336. [PubMed] [Google Scholar]
  33. Pan Y. T., Elbein A. D. Control of N-linked oligosaccharide synthesis: cellular levels of dolichyl phosphate are not the only regulatory factor. Biochemistry. 1990 Sep 4;29(35):8077–8084. doi: 10.1021/bi00487a013. [DOI] [PubMed] [Google Scholar]
  34. Patterson M. K., Jr Measurement of growth and viability of cells in culture. Methods Enzymol. 1979;58:141–152. doi: 10.1016/s0076-6879(79)58132-4. [DOI] [PubMed] [Google Scholar]
  35. Rossignol D. P., Lennarz W. J., Waechter C. J. Induction of phosphorylation of dolichol during embryonic development of the sea urchin. J Biol Chem. 1981 Oct 25;256(20):10538–10542. [PubMed] [Google Scholar]
  36. Rush J. S., Waechter C. J. Expression of dolichol-linked saccharide intermediate synthesis during the development of B lymphocytes. Glycobiology. 1991 Mar;1(2):229–235. doi: 10.1093/glycob/1.2.229. [DOI] [PubMed] [Google Scholar]
  37. Sagami H., Kurisaki A., Ogura K. Formation of dolichol from dehydrodolichol is catalyzed by NADPH-dependent reductase localized in microsomes of rat liver. J Biol Chem. 1993 May 15;268(14):10109–10113. [PubMed] [Google Scholar]
  38. Schmitt J. W., Elbein A. D. Inhibition of protein synthesis also inhibits synthesis of lipid-linked oligosaccharides. J Biol Chem. 1979 Dec 25;254(24):12291–12294. [PubMed] [Google Scholar]
  39. Sharma C., Radhakrishnamurthy B., Berenson G. S. Activation of dolichol-phosphate mannosyltransferase by dibutryl cyclic AMP in rat liver. Biochem Biophys Res Commun. 1988 Sep 15;155(2):615–621. doi: 10.1016/s0006-291x(88)80539-4. [DOI] [PubMed] [Google Scholar]
  40. Starr C. M., Lucas J. J. Regulation of dolichyl phosphate-mediated protein glycosylation: estrogen effects on glucosyl transfers in oviduct membranes. Arch Biochem Biophys. 1985 Feb 15;237(1):261–270. doi: 10.1016/0003-9861(85)90277-2. [DOI] [PubMed] [Google Scholar]
  41. Sumbilla C., Waechter C. J. Dolichol kinase, phosphatase, and esterase activity in calf brain. Methods Enzymol. 1985;111:471–482. doi: 10.1016/s0076-6879(85)11032-3. [DOI] [PubMed] [Google Scholar]
  42. Vijay I. K., Oka T. Developmental regulation of glycosyltransferases involved in biosynthesis of asparagine-linked glycoproteins in mouse mammary gland. Eur J Biochem. 1986 Jan 2;154(1):57–62. doi: 10.1111/j.1432-1033.1986.tb09358.x. [DOI] [PubMed] [Google Scholar]
  43. Welply J. K., Lau J. T., Lennarz W. J. Developmental regulation of glycosyltransferases involved in synthesis of N-linked glycoproteins in sea urchin embryos. Dev Biol. 1985 Jan;107(1):252–258. doi: 10.1016/0012-1606(85)90393-8. [DOI] [PubMed] [Google Scholar]
  44. White D. A., Speake B. K. The effect of cycloheximide on the glycosylation of lactating-rabbit mammary glycoproteins. Biochem J. 1980 Oct 15;192(1):297–301. doi: 10.1042/bj1920297. [DOI] [PMC free article] [PubMed] [Google Scholar]

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