Protein kinase-dependent overexpression of the nuclear protein pirin in c-JUN and RAS transformed fibroblasts.
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 alpha-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.
Bergman
A C
AC
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Alaiya
A A
AA
Wendler
W
W
Binetruy
B
B
Shoshan
M
M
Sakaguchi
K
K
Bergman
T
T
Kronenwett
U
U
Auer
G
G
Appella
E
E
Jörnvall
H
H
Linder
S
S
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000502:physiology; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D049308:Dioxygenases; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D016758:Genes, jun; D011905:Genes, ras; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D048369:MAP Kinase Kinase 1; D013058:Mass Spectrometry; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020929:Mitogen-Activated Protein Kinase Kinases; D020928:Mitogen-Activated Protein Kinases; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D009953:Ornithine-Oxo-Acid Transaminase / Q000096:biosynthesis / Q000235:genetics; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000096:biosynthesis / Q000235:genetics; D010735:Phosphoglycerate Kinase / Q000096:biosynthesis / Q000235:genetics; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D019908:Proto-Oncogene Proteins c-raf / Q000502:physiology; D051381:Rats; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics*; D014162:Transfection
D002352:Carrier Proteins; D005419:Flavonoids; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D009687:Nuclear Proteins; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D012261:Ribonucleoproteins; D049308:Dioxygenases; C105340:PIR protein, human; D009953:Ornithine-Oxo-Acid Transaminase; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D019908:Proto-Oncogene Proteins c-raf; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D048369:MAP Kinase Kinase 1; C482125:MAP2K1 protein, human; D020929:Mitogen-Activated Protein Kinase Kinases; D010735:Phosphoglycerate Kinase; D010751:Phosphopyruvate Hydratase; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one