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Cellular and Molecular Life Sciences CMLS

, Volume 55, Issue 3, pp 467–471 | Cite as

Protein kinase-dependent overexpression of the nuclear protein pirin in c-JUN and RAS transformed fibroblasts

  • A.-C. Bergman
  • A. A. Alaiya
  • W. Wendler
  • B. Binetruy
  • M. Shoshan
  • K. Sakaguchi
  • T. Bergman
  • U. Kronenwett
  • G. Auer
  • E. Appella
  • H. Jörnvall
  • S. Linder

Abstract.

Signalling via the protein kinase Raf-MEK-ERK pathway is of major importance for transformation by oncogenes. To identify genes affected by inhibition of this pathway, c-JUN transformed rat fibroblasts were treated with a MEK1 inhibitor (PD98059) and subjected to two-dimensional gel electrophoresis after cell lysis. Gene products with expression influenced by MEK1 inhibition were determined by mass spectrometry of fragments from in-gel tryptic digestions. The expression of pirin, a nuclear factor I-interacting protein, was lowered after inhibition of MEK1. Western blot analysis revealed increased expression of pirin in RAS and c-JUN transformed cells in the absence of PD98059. Inhibition of MEK1 also led to reduced expression of α-enolase, phosphoglycerate kinase, elongation factor 2 and heterogeneous nuclear ribonucleoprotein A3, the latter two being detected as truncated proteins. In contrast, the level of ornithine aminotransferase was increased. We conclude that inhibition of MEK1 results in major alterations of protein expression in c-JUN transformed cells, suggesting that this pathway is important for oncogene-induced phenotypic changes.

Key words. Two-dimensional gel electrophoresis; mass spectrometry; cell transformation. 

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Copyright information

© Birkhäuser Verlag Basel, 1999

Authors and Affiliations

  • A.-C. Bergman
    • 1
  • A. A. Alaiya
    • 2
  • W. Wendler
    • 3
  • B. Binetruy
    • 4
  • M. Shoshan
    • 5
  • K. Sakaguchi
    • 6
  • T. Bergman
    • 1
  • U. Kronenwett
    • 5
  • G. Auer
    • 2
  • E. Appella
    • 6
  • H. Jörnvall
    • 1
  • S. Linder
    • 5
  1. 1.Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm (Sweden)SE
  2. 2.Unit of Cell and Molecular Analysis, Department of Oncology and Pathology, Karolinska Hospital, S-171 76 Stockholm (Sweden)SE
  3. 3.Department of Biochemistry, Ludwig-Maximilian-University, Feodor-Lünen-Str. 25, D-81377 München (Germany)DE
  4. 4.Department of Cancer Research, National Center of Scientific Research, F-94801 Villejuif (France)FR
  5. 5.Radiumhemmet's Research Laboratory, Department of Oncology and Pathology, Karolinska Hospital, S-171 76 Stockholm (Sweden)SE
  6. 6.Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda (Maryland 20892, USA)US

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