Phosphorylation of histones is stimulated by phorbol esters in quiescent Reuber H35 hepatoma cells

A. P. Butler, C. V. Byus, Thomas J Slaga

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Abstract

Histones isolated from Reuber H35 rat hepatoma cells treated with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) were examined for possible alterations in phosphorylation. Incorporation of 32P orthophosphate into individual acid-extracted histones was monitored by autoradiography and scintillation counting of polyacrylamide gels or by reverse-phase high performance liquid chromatography. Treatment of quiescent H35 cells (arrested by serum starvation) with submicromolar doses of TPA resulted in a rapid and specific increase in phosphorylation of histones H2B and H10. Smaller increases in phosphorylation were observed for H4. No significant change in phosphorylation of the major H1 histones or H2A were observed after 1 h of treatment. The phosphorylation was TPA dose-dependent, with a maximum increase of approximately 14-fold for H2B, 11-fold for H10, and 2-fold for H4 achieved at 0.8 M TPA. The nonpromoting parent compound phorbol did not induce any of these changes. Furthermore, the mitogenic hormone insulin did not cause a similar pattern of histone phosphorylation, suggesting that the effect observed was not due to a general mitogenic response in the H35 hepatoma cells. Addition of 8-Br-cAMP also failed to reproduce the effect of TPA on histone phosphorylation, suggesting that cAMP-dependent protein kinases are not likely to be involved in mediating this response to TPA.

Original languageEnglish (US)
Pages (from-to)9421-9425
Number of pages5
JournalJournal of Biological Chemistry
Volume261
Issue number20
StatePublished - 1986
Externally publishedYes

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Phosphorylation
Phorbol Esters
Histones
Tetradecanoylphorbol Acetate
Hepatocellular Carcinoma
Acetates
Scintillation Counting
High performance liquid chromatography
Scintillation
Reverse-Phase Chromatography
Starvation
Cyclic AMP-Dependent Protein Kinases
Autoradiography
Carcinogens
Rats
Phosphates
High Pressure Liquid Chromatography
Hormones
Insulin
Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of histones is stimulated by phorbol esters in quiescent Reuber H35 hepatoma cells. / Butler, A. P.; Byus, C. V.; Slaga, Thomas J.

In: Journal of Biological Chemistry, Vol. 261, No. 20, 1986, p. 9421-9425.

Research output: Contribution to journalArticle

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AU - Byus, C. V.

AU - Slaga, Thomas J

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