Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells

Bernd J. Pulverer, Kenneth Hughes, Christopher C. Franklin, Andrew Kraft, Sally J. Leevers, James R. Woodgett

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Phorbol esters, such as phorbol myristate acetate (PMA), cause differentiation of U937 human monomvelocytic cells along the macrophage pathway. Within 15 min of PMA treatment DNA binding of the c-jun transcription factor is increased and is accompanied by rapid changes in the phosphate content of the c-jun protein. Phorbol esters stimulate phosphorylation of serines 63 and 73 located within the Al transactivation domain of c-Jun that have previously been shown to positively regulate activity. A protein kinase activity is detectable in extracts of phorbol ester-treated U937 cells that specifically targets these two serines. Using novel assays, the protein kinase activity has been purified over 1000-fold. The major portion of protein kinase activity co-chromatographs over three columns with pp42/44 mitogen-activated protein kinases as judged by immunological methods. The significance of these results with respect to mitogen-induced transcription of AP-1-responsive genes is discussed.

Original languageEnglish (US)
Pages (from-to)407-415
Number of pages9
JournalOncogene
Volume8
Issue number2
StatePublished - Feb 1993
Externally publishedYes

Fingerprint

U937 Cells
Phorbol Esters
Mitogen-Activated Protein Kinases
Protein Kinases
Phosphotransferases
Tetradecanoylphorbol Acetate
Serine
Proto-Oncogene Proteins c-jun
Transcription Factor AP-1
Mitogens
Transcriptional Activation
Transcription Factors
Macrophages
Phosphates
Phosphorylation
DNA
Genes

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Pulverer, B. J., Hughes, K., Franklin, C. C., Kraft, A., Leevers, S. J., & Woodgett, J. R. (1993). Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells. Oncogene, 8(2), 407-415.

Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells. / Pulverer, Bernd J.; Hughes, Kenneth; Franklin, Christopher C.; Kraft, Andrew; Leevers, Sally J.; Woodgett, James R.

In: Oncogene, Vol. 8, No. 2, 02.1993, p. 407-415.

Research output: Contribution to journalArticle

Pulverer, BJ, Hughes, K, Franklin, CC, Kraft, A, Leevers, SJ & Woodgett, JR 1993, 'Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells', Oncogene, vol. 8, no. 2, pp. 407-415.
Pulverer, Bernd J. ; Hughes, Kenneth ; Franklin, Christopher C. ; Kraft, Andrew ; Leevers, Sally J. ; Woodgett, James R. / Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells. In: Oncogene. 1993 ; Vol. 8, No. 2. pp. 407-415.
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