Skip to main content
SpringerLink
Log in

Incorporation of 2-14C-acetate into isolated nuclei and cytosolic lipids of rat thymus cells

  • Published:
Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Aims and scope

Abstract

It is generally recognized nowadays that active lipid metabolism takes place in the nucleus of a mammalian cell. Experimental data testify to the biosynthesis of polyphosphoinositides and phosphatidylcholine and reveal corresponding enzymes within nuclei of mammalian cells. These findings suggest that lipidmediated signaling pathways in nuclei operate independently of lipid-mediated regulatory mechanisms functioning in membranes and cytosol. To explore the pathways of intranuclear lipid biosynthesis, we studied incorporation of 2-14C-acetate into lipids of cytosol and isolated nuclei of rat thymus cells after separate and combined incubation with the labeled precursor. The most efficient incorporation of 2-14C-acetate into lipids (cholesterol, free fatty acids, and phospholipids) was observed in a reaction mixture containing cytosol. When the reaction mixture contained only nuclei, incorporation of the radioactive precursor into lipids also took place, but specific radioactivity of the lipids was essentially lower than in the cytosol. In both cases, 2-14C-acetate incorporated into phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol, and cardiolipin. Phosphatidylcholine, the most abundant membrane phospholipid, demonstrated the lowest radioactivity, which was significantly lower than that of phosphatidylethanolamine. Incorporation of newly synthesized free fatty acids in nuclear phospholipids was inhibited, if nuclei were incubated with cytosol. As a result, radioactive free fatty acids were accumulated in nuclei, while in cytosol they were efficiently incorporated into phospholipids. The levels of phospholipids and cholesterol remained constant regardless of incubation protocol, while the overall yield of free fatty acids decreased after combined incubation of nuclear and cytosolic fractions or after incubation of cytosol without nuclei. Putative mechanisms underlying the appearance of radioactive lipids in isolated nuclei of thymus cells are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Hunt, A.N., Completing the Cycles; The Dynamics of Endonuclear Lipidomics, Biochim. et Biophys. Acta, 2006, vol. 1761, nos. 5–6, pp. 577–587.

    Article  CAS  Google Scholar 

  2. Leeden, R.W. and Wu, G., Nuclear Lipids: Key Signaling Effectors in the Nervous System and Other Tissues, J. Lipid Res., 2004, vol. 45, no. 1, pp. 1–8.

    Article  Google Scholar 

  3. Martelli, A.M., Tabellini, G., Borgatti, P., Bortul, R., Capitani, S., and Neri, L.M., Nuclear Lipids: New Function for Old Molecules?, J. Cell. Biochem., 2003, vol. 88, no. 3, pp. 455–461.

    Article  Google Scholar 

  4. Irvine, R.F., Nuclear Lipid Signaling, Mol. Cell. Biol., 2003, vol. 4, no. 1, pp. 1–12.

    Google Scholar 

  5. Tamiya-Koizumi, K., Nuclear Lipid Metabolism and Signaling, J. Biochem., 2002, vol. 132, no. 1, pp. 13–22.

    Article  Google Scholar 

  6. Goppelt-Strube, M. and Resch, K., Polyunsaturated Fatty Acids Are Enriched in the Plasma Membranes of Mitogen-Stimulated T -Lymphocytes, Biochim. et Biophys. Acta, 1987, vol. 904, no. 1, pp. 22–28.

    Article  CAS  Google Scholar 

  7. Singh, J.K., Dasgupta, A., Adayev, T., Shahmehdi, S.A., Hammond, D., and Banerjee, P., Apoptosis Is Associated with an Increase in Saturated Fatty Acid Containing Phospholipids in the Neuronal Cell Line, HN2-5, Biochim. et Biophys. Acta, 1996, vol. 1304, no. 3, pp. 171–178.

    Article  CAS  Google Scholar 

  8. Tomassoni, M.-L., Amori, D., and Viola Magni, M., Changes of Nuclear Membrane Lipid Composition Affect RNA Nucleocytoplasmic Transport, Biochem. Biophys. Res. Commun., 1999, vol. 258, no. 2, pp. 476–481.

    Article  Google Scholar 

  9. Kolomiytseva, I.K., Kulagina, T.P., Markevich, L.N., Archipov, V.I., Slozhenikina, L.V., Fialkovskaya, L.A., and Potekhina, N.I., Nuclear and Chromatin Lipids: Metabolism in Normal and ?Irradiated Rats, Bioelectrochemistry, 2002, vol. 58, no. 1, pp. 31–39.

    Article  CAS  Google Scholar 

  10. Kulagina, T.P., Markevich, L.N., Kolomiytseva, I.K., and Alessenko, A.V., Synthesis of Lipids in Isolated Nuclei from Rat Thymus and Liver Cells, Biokhimiya (Rus.), 2003, vol. 68, no. 5, pp. 698–705.

    Google Scholar 

  11. Ves-Losada, A. and Brenner, R.R., Fatty Acid A5 Desaturation in Rat Liver Cell Nuclei, Mol. Cell. Biochem., 1995, vol. 142, no. 2, pp. 163–170.

    Article  CAS  Google Scholar 

  12. Baker, R.R. and Chang, H.-Y., The Rapid Incorporation of Radioactive Fatty Acid into Triacylglycerols during the In vitro Acylation of Native Lipids of Neuronal Nuclei, Biochim. et Biophys. Acta, 1983, vol. 752, no. 1, pp. 1–9.

    Article  CAS  Google Scholar 

  13. Allfrey, V.G., Biosynthetic Reactions in the Cell Nucleus, Aspects of Protein Biosynthesis, Anfinsen, C.B., Jr., Ed., N.Y.-London: Acad. Press, 1970, pp. 247–365.

  14. Gerlach, I. and Deuticke, B., Eine Einfache Metode zur Microbestimmung den Phosphate in der Papir Chromatographie, Biochemische Z., 1963, vol. 337, no. 5, pp. 477–482.

    CAS  Google Scholar 

  15. Marsh, J.B. and Weinstein, D.B., Simple Charring Method for Determination of Lipids, J. Lipid Res., 1966, vol. 7, no. 4, pp. 574–576.

    Article  CAS  Google Scholar 

  16. Molecular Cloning. A Laboratory Manual, Sambrook, J., Fritsch, E.F., and Maniatis, T., Eds., N.Y: Cold Spring Harbor Laboratory, 1989, vol. 1, pp. 6.9–6.19.

  17. Biardi, L. and Krisans, S.K., Compartmentalization of Cholesterol Biosynthesis. Conversion of Mevalonate to Farnesyl Diphosphate Occurs in the Peroxisomes, J. Biol. Chem., 1996, vol. 271, no. 3, pp. 1784–1788.

    Article  CAS  Google Scholar 

  18. Hogenboom, S., Wanders, R.J., and Waterham, H.R., Cholesterol Biosynthesis Is Not Defective in Peroxisome Biogenesis Defective Fibroblasts, Mol. Genet. Metab., 2003, vol. 80, no. 3, pp. 290–295.

    Article  CAS  Google Scholar 

  19. Kulagina, T.P., Kolomiytseva, I.K., Moiseeva, S.A., and Kuzin, A.M., Non-Monotonous Changes in Lipid Metabolism of Thymus Cell Nuclei under Chronic Exposure of Rats to 7-Irradiation with the Dose 3 cGy/Day, Doklady RAN (Rus.), 2000, vol. 370, no. 3, pp. 403–406.

    CAS  Google Scholar 

  20. Kulagina, T.P., Shuruta, S.A., Kolomiytseva, I.K., and Vakulova, L.A., The Action of Continuous γ-Irradiation at a Low-Dose Rate and β-Carotene on Lipid Metabo lism of Rat Thymus Nuclei, Bulleten' Eksperimental'noi Biologii i Meditsiny (Rus.), 1998, vol. 126, no. 9, pp. 311–313.

    CAS  Google Scholar 

  21. Cullis, P.R. and De Kruijff, B., Lipid Polymorphism and the Functional Roles of Lipids in Biological Membranes, Biochim. et Biophys. Acta, 1979, vol. 559, no. 4, pp. 399–420.

    Article  CAS  Google Scholar 

  22. De Kruijff, B., Lipid Polymorphism and Biomembrane Function, Curr Opin. Chem. Biol., 1997, vol. 1, no. 4, pp. 564–569.

    Article  Google Scholar 

  23. Mikhaleva, N.I., Golovastov, V.V., Zolov, S.N., Bogdanov, M.V., Dowhan, W., and Nesmeyanova, M.A., Depletion of Phosphatidylethanolamine Affects Secre tion of Escherichia coli Alkaline Phosphatase and Its Transcriptional Expression, FEES Lett., 2001, vol. 493, no. 2–3, pp. 85–90.

    Article  CAS  Google Scholar 

  24. Golovastov, V.V., Zolov, S.N., and Nesmeyanova, M.A., Study of Interaction of Export Initiation Domain of Escherichia coli Mature Alkaline Phosphatase with Membrane Phospholipids during Secretion, Biokhimiya (Rus.), 2002, vol. 67, no. 9, pp. 1182–1190.

    Google Scholar 

  25. Lazebnik, Y.A., Cole, S., Cooke, C.A., Nelson, W.G., and Earnshaw, W.C., Nuclear Events of Apoptosis In vitro in Cell-Free Mitotic Extracts: A Model System for Analysis of the Active Phase of Apoptosis, J. Cell. Biol., 1993, vol. 123, no. 1, pp. 7–22.

    Article  CAS  Google Scholar 

  26. Martin, S.J., Newmeyer, D.D., Mathias, S., Farschon, D.M., Wang, H.-G., Reed, J.C., Kolesnick, R.N., and Green, D.R., Cell-Free Reconstitution of Fas-, UV-Radiation- and Ceramide-Induced Apoptosis, EMBO J., 1995, vol. 14, no. 21, pp. 5191–5200.

    Article  CAS  Google Scholar 

  27. Enari, M., Hase, A., and Nagata, S., Apoptosis by a Cytotosolic Extract from Fas-Activated Cells, EMBO J., 1995, vol. 14, no. 21, pp. 5201–5208.

    Article  CAS  Google Scholar 

  28. Zhao, Y., Wu, M., Shen, Y., and Zhai, Z., Analysis of Nuclear Apoptotic Process in a Cell-Free System, Cell. Mol. Life Sci., 2001, vol. 58, no. 2, pp. 298–306.

    Article  CAS  Google Scholar 

  29. Nikonova, L.V., Nelipovich, P.A., and Umansky, S.R., The Involvement of Nuclear Nucleases in Rat Thymocyte DNA Degradation after 7l Irradiation, Biochim. et Biophys. Acta, 1982, vol. 699, no. 3, pp. 281–289.

    Article  CAS  Google Scholar 

  30. Ves-Losada, A., Mate, S.M., and Brenner, R.R., Incorporation and Distribution of Saturated and Unsaturated Fatty Acids into Nuclear Lipids of Hepatic Cells, Lipids, 2001, vol. 36, no. 3, pp. 273–282.

    Article  CAS  Google Scholar 

  31. Glatz, J.F.C. and Van Der Vusse, G.J., Cellular Fatty Acid-Binding Proteins: Their Function and Physiological Significance, Progr Lipid Res., 1996, vol. 35, no. 3, pp. 243–282.

    Article  CAS  Google Scholar 

  32. Schmelter, T., Trigatti, B.L., Gerber, G.E., and Mangroo, D., Biochemical Demonstration of the Involvement of Fatty Acyl-CoA Synthetase in Fatty Acid Trans location across the Plasma Membrane, J. Biol. Chem., 2004, vol. 279, no. 23, pp. 24163–24170.

    Article  CAS  Google Scholar 

  33. Ehehalt, R., Fullekrug, J., Pohl, J., Ring, A., Herrman, T., and Stremmel, W., Translocation of Long Chain Fatty Acids across the Plasma Membrane-Lipid Rafts and Fatty Acid Transport Proteins, Mol. Cell. Biochem., 2006, vol. 284, nos. 1–2, pp. 135–140.

    Article  CAS  Google Scholar 

  34. Meshulam, T., Simard, J.R., Wharton, J., Hamilton, J.A., and Pilch, PF., Role of Caveolin-1 and Cholesterol in Transmembrane Fatty Acid Movement, Biochemistry, 2006, vol. 45, no. 9, pp. 2882–2893.

    Article  CAS  Google Scholar 

  35. Marmunti, M. and Catala, A., Incorporation of 1-14C-Linoleic Acid in Rat Liver Nuclei and Chromatin Fractions, Biochem. Cell Biol., 2001, vol. 33, no. 3, pp. 261–267.

    Article  CAS  Google Scholar 

  36. Rung, E., Friberg, P.A., Shao, R., Larsson, D.G.J., Nielson, E.Ch., Svensson, P.-A., Carlsson, B., Carlsson, L.M.S., and Billig, H., Progesterone-Receptor Antagonists and Statins Decrease De novo Cholesterol Synthesis and Increase Apoptosis in Rat and Human Periovulatory Granulose Cells In vitro, Biol. Reprod., 2005, vol. 72, no. 3, pp. 538–545.

    Article  CAS  Google Scholar 

  37. Nakamura, M.T., Cheon, Y., Li, Y., and Nara, T.Y., Mechanisms of Regulation of Gene Expression by Fatty Acids, Lipids, 2004, vol. 39, no. 11, pp. 1077–1083.

    Article  CAS  Google Scholar 

  38. Murthy, S., Tong, H., and Hohl, R.J., Regulation of Fatty Acid Synthesis by Farnesyl Pyrophosphate, J. Biol. Chem., 2005, vol. 280, no. 51, pp. 41793–41804.

    Article  CAS  Google Scholar 

  39. Kuzin, A.M., Molekulyarnaya radiobiologiya kletochnogo yadra (Molecular Radiobiology of Cell Nucleus), Moscow: Atomizdat, 1973.

    Google Scholar 

  40. Wanders, R.J., Peroxisomes, Lipid Metabolism and Peroxisomal Disorders, Mol. Genet. Metab., 2004, vol. 83, nos. 1–2, pp. 16–27.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Cell Biophysics, Russian Academy of Sciences, 142290, Pushchino, Moscow oblast, Russia

    T. P. Kulagina

Authors
  1. T. P. Kulagina
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. P. Kulagina.

Additional information

Original Russian Text © T.P. Kulagina, 2008, published in Biologicheskie Membrany, 2008, Vol. 25, No. 1, pp. 50–57.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kulagina, T.P. Incorporation of 2-14C-acetate into isolated nuclei and cytosolic lipids of rat thymus cells. Biochem. Moscow Suppl. Ser. A 2, 48–54 (2008). https://doi.org/10.1134/S199074780801008X

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S199074780801008X

Keywords

Search

Navigation