Abstract
Primary cultures of neonatal cardiac myocytes were used to determine the effects of tumor-promoting phorbol esters on ribosomal RNA (rRNA) synthesis during myocyte growth. Treatment of myocytes with phorbol-12,13-dibutyrate (PDBu) increased protein accumulation by 25% and RNA content by 20%. Rates of rRNA synthesis were measured to assess the mechanism by which rRNA accumulated during myocyte growth. Rates of rRNA synthesis were determined from the incorporation of [3H]uridine into UMP of purified rRNA and the specific radioactivity of the cellular UTP pool. After 24h of PDBu treatment, cellular rates of 18S and 28S rRNA synthesis were accelerated by 67% and 64%, respectively. The increased rate of rRNA synthesis accounted for the net increase in myocyte rRNA content after PDBu treatment.
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Morgan HE, Gordon EE, Kira Y, Chua BHL, Russo LA, Peterson CJ, McDermott PJ, Watson PA: Biochemical mechanisms of cardiac hypertrophy. Ann Rev Physiol 49: 533–543, 1987
Young VR: The role of skeletal and cardiac muscle in the regulation of protein metabolism. In: HN Munro and JB Allison (eds) Mammalian Protein Metabolism, Academic Press, New York, 1970, pp 585–662
Siehl D, Shua BHL, Lautensach-Belser N, Morgan HE: Faster protein and ribosome synthesis in thyroxine-induced hypertrophy of rat heart. Am J Physiol 248: C309-C319, 1985
McDermott PJ, Morgan HE: Contraction modulates the capacity for protein synthesis during growth of neonatal heart cells in culture. Cite Res 64: 542–553, 1989
Sollner-Webb B, Tower J: Transcription of cloned eukaryotic ribosomal RNA genes. Ann Rev Biochem 55: 801–830, 1986
Nishizuka Y: The role of protein kinase C in cell surface signal transduction and tumor production. Nature Lond 308: 693–698, 1984
Kikkawa V, Takai Y, Tanaka Y, Miyake R, Nishizuka Y: Protein kinase C as a possible receptor protein of tumor-promoting phorbol esters. J Biol Chem 258: 11442–11445, 1983
Henrich CJ, Simpson PC: Differential acute and chronic response of protein kinase C in cultured neonatal rat heart myocytes to al-adrenergic and phorbol ester stimulation. J Mol Cell Cardiol20: 1081–1085, 1988
McDermott PJ, Rothblum LL Smith SD, Morgan HE: Accelerated rates of ribosomal RNA synthesis during growth of contracting heart cells in culture. J Biol Chem 264: 18220–18227, 1989
Lowry DH, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193: 265–275, 1951
Cesarone CF, Bolognesi C, Santi L: Improved microfluorometric DNA determination in biological material using 33258 Hoechst. Anal Biochem 100: 188–197, 1979
Folley LA, Power SC, Poyton RO: Separation of nucleotides by ion-pair, reversed-phase high performance liquid chromatography. J Chromatogr 281: 199–207, 1983
Khym JX: An analytical system for rapid separation of tissue nucleotides at low pressures on conventional anion exchangers. Clin Chem 21: 1245–1252, 1975
Matsushita S, Fanburg BL: Pyrimidine nucleotide synthesis in the normal and hypertrophying rat heart. Cite Res 27: 415–428, 1970
Chan Y, Gutell R, Noller HF, Wool IG: The nucleotide sequence of a rat 18S ribosomal ribonucleic acid gene and a proposal for the secondary structure of 188 ribosomal ribonucleic acid. J Biol Chem 259: 224–230, 1984
Chan Y, Olvera J, Wool IG: The structure of rat 288 ribosomal ribonucleic acid inferred from the sequence of nucleotides in a gene. Nucleic Acids Res 11: 7819–7831, 1983
Ashford AJ, Pain VM: Effect of diabetes on the rates of synthesis and degradation of ribosomes in rat muscle and liver in vivo. J Biol Chem 261: 4059–4065, 1986
Limas CJ: Characterization of phorbol ester binding to isolated cardiac myocytes. Arch Biochem Biophys 238: 300–304, 1985
Yuan S, Sunahara FA, Sen AK: Tumor-promoting phorbol esters inhibit cardiac functions and induce redistribution of protein kinase C in perfused beating rat heart. Cite Res 61: 372–378, 1987
Gertz BJ, Haugaard ES, Haugaard N: Effects of thyroid hormone on UTP content and uridine kinase activity of rat heart and skeletal muscle. Am J Physiol 238: E443-E449, 1980
Jacobs FA, Bird RC, Sells BH: Differentiation of rat myoblasts. Regulation of turnover of ribosomal proteins and their mRNAs. Eur J Biochem 150: 255–263, 1985
Hammond ML, Bowman LH: Insulin stimulates the translation of ribosomal proteins and the transcription of rDNA in mouse myoblasts. J Biol Chem 263: 17785–17791, 1988
Wolf S, Sameshima M, Liebhaber SA, Schlessinger D: Regulation of ribosomal ribonucleic acid levels in growing, 3H-arrested, and crisis-phase WI-38 human diploid fibroblasts. Biochemistry 19: 3484–3490, 1980
Angel P, Imagawa M, Chiu R, Stein B, Imbra RJ, Rahmsdorf HJ, Jonat C, Herrlich P, Karin M: Phobol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell 49: 729–739, 1987
Lee W, Mitchell P, Tjian R: Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell 49: 741–752, 1987
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Hanedal, T., McDermott, P.J. Stimulation of ribosomal RNA synthesis during hypertrophic growth of cultured heart cells by phorbol ester. Mol Cell Biochem 104, 169–177 (1991). https://doi.org/10.1007/BF00229817
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DOI: https://doi.org/10.1007/BF00229817