Summary
Ornithine decarboxylase, a key enzyme in polyamine biosynthesis and cell growth, has been localized in mouse kidney by autoradiography after administration of radiolabeled α-difluoromethylornithine. This drug is an enzyme-activated irreversible inhibitor of ornithine decarboxylase and forms a covalent bond with the enzyme. It was found that ornithine decarboxylase is present in all cell types studied but that the highest content occurs in the proximal convoluted tubules followed by the distal convoluted tubules and the collecting tubules. The majority of the enzyme is located in the cytoplasm but about 10–15% is present in the nuclei (often associated with nucleolus-like components) of the cells of the proximal and distal convoluted tubules. The labeled ornithine decarboxylase was lost rapidly from both nucleus and cytoplasm of all the cell types examined, and labeling by radioactive α-difluoromethylornithine was greatly reduced if the mice were pretreated for 5 h with cycloheximide to block protein synthesis. These results indicate that ornithine decarboxylase turns over rapidly in all of the cells.
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References
Atmar VJ, Kuehn GD (1981) Phosphorylation of ornithine decarboxylase by a polyamine-dependent protein kinase. Proc Natl Acad Sci 78:5518–5522
Bartholeyns J (1983) Subcellular distribution of ornithine decarboxylase in rat liver and accessibility of the enzyme to α-difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase. Life Sci 32:1305–1312
Bitonti AJ, Couri D (1981) Drug-induced migration of cytoplasmic ornithine decarboxylase into rat liver nucleus is not related to increased RNA polymerase activity. Biochem Biophys Res Commun 99:1040–1044
Emanuelsson H, Heby O (1982) Ornithine decarboxylase activity in nucleolus and nucleoplasm demonstrated autoradiographically with tritium-labeled α-difluoromethylornithine. Cell Biol Int Rep 6:951–954
Erwin BG, Seely JE, Pegg AE (1983) Mechanism of stimulation of ornithine decarboxylase activity in transformed mouse fibroblasts. Biochemistry 22:3027–3032
Grillo MA, Fossa T (1983) Ornithine decarboxylase in rat liver nuclei. Int J Biochem 15:139–141
Grove J, Fozard JR, Mamont PS (1981) Assay of α-difluoromethylornithine in body fluids and tissues by automatic amino acid analysis. J Chromatogr 223:409–416
Heby O (1981) Role of polyamines in the control of cell proliferation and differentiation. Differentiation 14:1–20
Henningson A-C, Henningson S, Tjälve T, Hammar L, Löwendahl G (1983) Whole body and microautographic localization of ornithine decarboxylase in the kidneys of nandrolone-treated mice using tritium-labeled α-difluoromethylornithine. Adv Polyamine Res 4:719–726
Jänne J, Pösö H, Raina A (1978) Polyamines in rapid growth and cancer. Biochim Biophys Acta 473:241–293
McCormick F (1977) Polyamine metabolism in enucleated mouse L-cells. J Cell Physiol 93:285–2982
Metcalf BW, Bey P, Danzin C, Jung MJ, Casara P, Vevert JP (1978) Catalytic irreversible inhibition of mammalian ornithine decarboxylase (E.C. 4.1.1.17) by substrate and product analogoues. J Am Chem Soc 100:2551–2553
Murphy BJ, Brosnan ME (1976) Subcellular localization of ornithine decarboxylase in liver of control and growth hormone treated rats. Biochem J 157:33–39
Pegg AE, McCann PP (1982) Polyamine metabolism and function. Am J Physiol 243:C212-C221
Pegg AE, Williams-Ashman HG (1981) Biosynthesis of putrescine. In: Morris DR, Marton LJ (eds) Polyamines in biology and medicine. Marton New York: Dekker p 3–42
Pegg AE, Seely JE, Pösö H, Della-Ragione F, Zagon IS (1982a) Polyamine biosynthesis and interconversion in rodent tissue. Fed Proc 41:3065–3072
Pegg AE, Seely JE, Zagon IS (1982b) Autoradiographic localization of ornithine decarboxylase in mouse kidney using [5-14C]αdifluoromethylornithine. Science 217:68–70
Persson L, Rosengren E, Sundler F (1982) Localization of ornithine decarboxylase by immunocytochemistry. Biochem Biophys Res Commun 104:1196–1201
Persson L, Rosengren E, Sundler F, Uddman R (1983) Immunocytochemical demonstration of ornithine decarboxylase. Adv Polyamine Res 4:571–584
Pritchard ML, Seely JE, Pösö H, Jefferson LS, Pegg AE (1981) Binding of radioactive α-difluoromethylornithine to rat liver ornithine decarboxylase. Biochem Biophys Res Commun 100:1597–1603
Raina A, Jänne J (1975) Physiology of the natural polyamines, putrescine, spermidine and spermine. Med Biol 53:121–147
Russell DH (1980) Ornithine decarboxylase as a biological and pharmacological tool. Pharmacology 20:117–129
Russell DH (1981) Ornithine decarboxylase: a key regulatory protein. Med Biol 59:286–295
Seely JE, Pösö H, Pegg AE (1982a) Effect of androgens on turnover of ornithine decarboxylase in mouse kidney studies using labeling of the enzyme by reaction with [14C]α-difluoromethylornithine. J Biol Chem 257:7549–7553
Seely JE, Pösö H, Pegg AE (1982b) Measurement of the number of ornithine decarboxylase molecules in rat and mouse tissues under various physiological conditions by binding of radiolabeled α-difluoromethylornithine. Biochem J 206:311–318
Snyder SH, Kreuz DS, Meding VJ, Russell DH (1970) Polyamine synthesis and turnover in rapidly growing tissues. Ann NY Acad Sci 171:749–771
Winer (1971) Statistical principles in experimental design. 2nd Edition McGraw-Hill, New York, pp 191–195
Zagon IS, Seely JE, Pegg AE (1983) Autoradiographic localization of ornithine decarboxylase. Methods Enzymol 94:169–176
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Zagon, I.S., McLaughlin1, P.J., Seely, J.E. et al. Autoradiographic localization of ornithine decarboxylase in mouse kidney by use of radiolabeled α-difluoromethylornithine. Cell Tissue Res. 235, 371–377 (1984). https://doi.org/10.1007/BF00217862
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DOI: https://doi.org/10.1007/BF00217862