Platelet-derived growth factor stimulates Src-dependent mRNA stabilization of specific early genes in fibroblasts

Paul A. Bromann, Hasan Korkaya, Craig P. Webb, Jeremy Miller, Tammy L. Calvin, Sara A. Courtneidge

Research output: Contribution to journalArticle

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Abstract

The Src family of protein-tyrosine kinases (SFKs) participates in a variety of signal transduction pathways, including promotion of cell growth, prevention of apoptosis, and regulation of cell interactions and motility. In particular, SFKs are required for the mitogenic response to platelet-derived growth factor (PDGF). However, it is not clear whether there is a discrete SFK-specific pathway leading to enhanced gene expression or whether SFKs act to generally enhance PDGF-stimulated gene expression. To examine this, we treated quiescent NIH3T3 cells with PDGF in the presence or absence of small molecule inhibitors of SFKs, phosphatidylinositol 3-kinase (PI3K), and MEK1/2. Global patterns of gene expression were analyzed by using Affymetrix Gene-Chip arrays, and data were validated by using reverse transcription-PCR and ribonuclease protection assay. We identified a discrete set of immediate early genes induced by PDGF and inhibited in the presence of the SFK-selective inhibitor SU6656. A subset of these SFK-dependent genes was induced by PDGF even in the presence of the MEK1/2 inhibitor U0126 or the PI3K inhibitor LY294002. By using ribonuclease protection assays and nuclear run-off assays, we further determined that PDGF did not stimulate the rate of transcription of these SFK-dependent immediate early genes but rather promoted mRNA stabilization. Our data suggest that PDGF regulates gene expression through an SFK-speeific pathway that is distinct from the Ras-MAPK and PI3K pathways, and that SFKs signal gene expression by enhancing mRNA stability.

Original languageEnglish (US)
Pages (from-to)10253-10263
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number11
DOIs
StatePublished - Mar 18 2005

Fingerprint

src-Family Kinases
Platelet-Derived Growth Factor
Fibroblasts
Protein-Tyrosine Kinases
Stabilization
Genes
Messenger RNA
Gene expression
Phosphatidylinositol 3-Kinase
Assays
Transcription
Ribonucleases
Gene Expression
Signal transduction
Immediate-Early Genes
Cell growth
Cells
Apoptosis
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Platelet-derived growth factor stimulates Src-dependent mRNA stabilization of specific early genes in fibroblasts. / Bromann, Paul A.; Korkaya, Hasan; Webb, Craig P.; Miller, Jeremy; Calvin, Tammy L.; Courtneidge, Sara A.

In: Journal of Biological Chemistry, Vol. 280, No. 11, 18.03.2005, p. 10253-10263.

Research output: Contribution to journalArticle

Bromann, Paul A. ; Korkaya, Hasan ; Webb, Craig P. ; Miller, Jeremy ; Calvin, Tammy L. ; Courtneidge, Sara A. / Platelet-derived growth factor stimulates Src-dependent mRNA stabilization of specific early genes in fibroblasts. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 11. pp. 10253-10263.
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