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  • Title: Epidermal growth factor induces c-fos and c-jun mRNA via Raf-1/MEK1/ERK-dependent and -independent pathways in bovine luteal cells.
    Author: Chen DB, Davis JS.
    Journal: Mol Cell Endocrinol; 2003 Feb 28; 200(1-2):141-54. PubMed ID: 12644307.
    Abstract:
    Epidermal growth factor (EGF) modulates the actions of gonadotropins in the corpus luteum. The membrane-associated EGF receptors undergo rapid tyrosine phosphorylation and internalization upon ligand binding in ovarian cells, including luteal cells. However, little is known about the post-receptor signaling events induced by EGF that lead to the transcriptional regulation of EGF-responsive genes in the ovary. The present study was designed to examine in bovine luteal cells (1) activation of the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) signaling cascade (Raf/MEK/ERK) by EGF; (2) mRNA expression of AP-1 transcription factors, i.e. c-fos and c-jun, in response to EGF; and (3) the role of ERK in EGF-induced expression of c-fos and c-jun mRNA. Raf-1 and B-Raf, but not A-Raf, were activated by EGF (10 ng/ml) and the pharmacological protein kinase C (PKC) activator phorbol myristate acetate (PMA, 20 nM). Activation of Raf resulted in the phosphorylation and activation of MAPK kinase (MEK1) which subsequently activated ERKs. Treatment with EGF-induced the phosphorylation of both ERK2 and ERK1 in a time and concentration dependent manner. Additionally, activated ERK was found in the nucleus of the cells following treatment with EGF (10 ng/ml) and PMA (PMA, 20 nM) for 5 min. Depletion of PKC by chronic PMA treatment (2.5 microM, 24 h) only partially inhibited the stimulatory effects of EGF on Raf-1, ERK2 and ERK1. These data demonstrate that PKC-dependent and independent-mechanisms are involved in EGF activation of the Raf/MEK/ERK signaling cascade in bovine luteal cells. EGF rapidly and transiently stimulated the expression of c-fos and c-jun mRNA in bovine luteal cells. Maximal induction of c-fos and c-jun mRNA by EGF occurred within 30 min of treatment with 10 ng/ml EGF. Treatment with the MEK1 inhibitor PD098059 (50 microM) abolished EGF-induced ERK activation. However, blocking EGF-induced ERK activation by pretreatment with PD098059 only partially attenuated EGF-induced c-fos and c-jun mRNA expression. Thus, additional pathways are implicated in the regulation of c-fos and c-jun mRNA expression by EGF in bovine luteal cells.
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