The transcriptional response to Raf activation is almost completely dependent on Mitogen-activated Protein Kinase Kinase activity and shows a major autocrine component.

The Raf protein kinases are major effectors of Ras GTPases and key components of the transcriptional response to serum factors, acting at least in part through the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway. It has recently been suggested that Raf also may trigger other as yet uncharacterized signaling pathways. Here, we have used cDNA microarrays to dissect changes in gene expression induced by activation of inducible c-Raf-1 constructs in human mammary epithelial and ovarian epithelial cells. The majority of Raf-induced transcriptional responses are shown to be blocked by pharmacological inhibition of the Raf substrate mitogen-activated protein kinase kinase, indicating that potential mitogen-activated protein kinase kinase-independent Raf signaling pathways have no significant influence on gene expression. In addition, we used epidermal growth factor receptor inhibitory drugs to address the contribution of autocrine signaling by Raf-induced EGF family proteins to the Raf transcriptional response. At least one-half of the transcription induced by Raf activation requires epidermal growth factor (EGF) receptor function The EGF receptor-independent component of the Raf transcriptional response is entirely up-regulation of gene expression, whereas the EGF receptor-dependent component is an equal mixture of up- and down-regulation. The use of transcriptional profiling in this way allows detailed analysis of the architecture of signaling pathways to be undertaken.

[1]  C. Der,et al.  Raf-independent Deregulation of p38 and JNK Mitogen-activated Protein Kinases Are Critical for Ras Transformation* , 2002, The Journal of Biological Chemistry.

[2]  D. Dowbenko,et al.  The Forkhead Transcription Factor AFX Activates Apoptosis by Induction of the BCL-6 Transcriptional Repressor* , 2002, The Journal of Biological Chemistry.

[3]  J. Sebolt-Leopold,et al.  Unraveling the complexities of the Raf/MAP kinase pathway for pharmacological intervention. , 2002, Trends in molecular medicine.

[4]  Jing Chen,et al.  Raf-1 promotes cell survival by antagonizing apoptosis signal-regulating kinase 1 through a MEK–ERK independent mechanism , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[5]  E. Wagner,et al.  Embryonic lethality and fetal liver apoptosis in mice lacking the c‐raf‐1 gene , 2001, The EMBO journal.

[6]  C. Pritchard,et al.  MEK kinase activity is not necessary for Raf‐1 function , 2001, The EMBO journal.

[7]  A. Schulze,et al.  Analysis of the transcriptional program induced by Raf in epithelial cells , 2001, Nature Genetics.

[8]  P. Lipsky,et al.  Raf-induced transformation requires an interleukin 1 autocrine loop. , 2001, Cancer research.

[9]  M. White,et al.  Uncoupling Raf1 from MEK1/2 Impairs Only a Subset of Cellular Responses to Raf Activation* , 2000, The Journal of Biological Chemistry.

[10]  W. Kolch Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. , 2000, The Biochemical journal.

[11]  A. Schulze,et al.  Raf induces TGFbeta production while blocking its apoptotic but not invasive responses: a mechanism leading to increased malignancy in epithelial cells. , 2000, Genes & development.

[12]  Fiona M. Watt,et al.  The EGF Receptor Provides an Essential Survival Signal for SOS-Dependent Skin Tumor Development , 2000, Cell.

[13]  T. Wirth,et al.  Raf induces NF-kappaB by membrane shuttle kinase MEKK1, a signaling pathway critical for transformation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Pouysségur,et al.  The p42/p44 MAP kinase pathway prevents apoptosis induced by anchorage and serum removal. , 2000, Molecular biology of the cell.

[15]  S. Yonehara,et al.  Oncogenic K-Ras and Basic Fibroblast Growth Factor Prevent FAS-Mediated Apoptosis in Fibroblasts through Activation of Mitogen-Activated Protein Kinase , 2000, The Journal of cell biology.

[16]  C. Marshall,et al.  How do small GTPase signal transduction pathways regulate cell cycle entry? , 1999, Current opinion in cell biology.

[17]  S. R. Datta,et al.  Cell survival promoted by the Ras-MAPK signaling pathway by transcription-dependent and -independent mechanisms. , 1999, Science.

[18]  E. Nishida,et al.  Activation of the Protein Kinase ERK5/BMK1 by Receptor Tyrosine Kinases , 1999, The Journal of Biological Chemistry.

[19]  C. Marshall,et al.  Activation of mitogen-activated protein kinase is necessary but not sufficient for proliferation of human thyroid epithelial cells induced by mutant Ras , 1999, Oncogene.

[20]  G. Cooper,et al.  B-Raf Inhibits Programmed Cell Death Downstream of Cytochrome c Release from Mitochondria by Activating the MEK/Erk Pathway , 1999, Molecular and Cellular Biology.

[21]  K. Guan,et al.  Negative Regulation of the Forkhead Transcription Factor FKHR by Akt* , 1999, The Journal of Biological Chemistry.

[22]  M. Sliwkowski,et al.  Binding specificities and affinities of egf domains for ErbB receptors , 1999, FEBS letters.

[23]  H. Pahl,et al.  Activators and target genes of Rel/NF-kappaB transcription factors. , 1999, Oncogene.

[24]  W. Denny,et al.  Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Troppmair,et al.  Activation of NF-κB by oncogenic Raf in HEK 293 cells occurs through autocrine recruitment of the stress kinase cascade , 1998, Oncogene.

[26]  F. Hobbs,et al.  Identification of a Novel Inhibitor of Mitogen-activated Protein Kinase Kinase* , 1998, The Journal of Biological Chemistry.

[27]  J Downward,et al.  Ras signalling and apoptosis. , 1998, Current opinion in genetics & development.

[28]  C. Der,et al.  Oncogenic Ha-Ras-induced Signaling Activates NF-κB Transcriptional Activity, Which Is Required for Cellular Transformation* , 1997, The Journal of Biological Chemistry.

[29]  E. Lees,et al.  Raf-induced proliferation or cell cycle arrest is determined by the level of Raf activity with arrest mediated by p21Cip1 , 1997, Molecular and cellular biology.

[30]  M. McMahon,et al.  Mutations of critical amino acids affect the biological and biochemical properties of oncogenic A-Raf and Raf-1 , 1997, Oncogene.

[31]  Minoru Takagi,et al.  Induction of Apoptosis by ASK1, a Mammalian MAPKKK That Activates SAPK/JNK and p38 Signaling Pathways , 1997, Science.

[32]  K. Diener,et al.  Molecular Cloning and Characterization of a Novel Protein Kinase with a Catalytic Domain Homologous to Mitogen-activated Protein Kinase Kinase Kinase* , 1996, The Journal of Biological Chemistry.

[33]  John Calvin Reed,et al.  Bcl-2 Targets the Protein Kinase Raf-1 to Mitochondria , 1996, Cell.

[34]  S. Oldham,et al.  Activation of the Raf-1/MAP kinase cascade is not sufficient for Ras transformation of RIE-1 epithelial cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[35]  R. Treisman,et al.  Regulation of transcription by MAP kinase cascades. , 1996, Current opinion in cell biology.

[36]  C. Marshall,et al.  Control of the ERK MAP kinase cascade by Ras and Raf. , 1996, Cancer surveys.

[37]  C. Marshall Ras effectors. , 1996, Current opinion in cell biology.

[38]  Philip R. Cohen,et al.  PD 098059 Is a Specific Inhibitor of the Activation of Mitogen-activated Protein Kinase Kinase in Vitro and in Vivo(*) , 1995, The Journal of Biological Chemistry.

[39]  J. Abraham,et al.  Rapid induction of heparin-binding epidermal growth factor/diphtheria toxin receptor expression by Raf and Ras oncogenes. , 1995, Genes & development.

[40]  A. Bridges,et al.  A synthetic inhibitor of the mitogen-activated protein kinase cascade. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[41]  M. Klagsbrun,et al.  Phorbol ester induces the rapid processing of cell surface heparin-binding EGF-like growth factor: conversion from juxtacrine to paracrine growth factor activity. , 1995, Molecular biology of the cell.

[42]  R. Berkowitz,et al.  Characterization of human ovarian surface epithelial cells immortalized by human papilloma viral oncogenes (HPV-E6E7 ORFs). , 1995, Experimental cell research.

[43]  M. Jaye,et al.  Activation of Cytosolic Phospholipase A by Basic Fibroblast Growth Factor via a p42 Mitogen-activated Protein Kinase-dependent Phosphorylation Pathway in Endothelial Cells (*) , 1995, The Journal of Biological Chemistry.

[44]  Sally J. Leevers,et al.  Requirement for Ras in Raf activation is overcome by targeting Raf to the plasma membrane , 1994, Nature.

[45]  D. Morrison,et al.  Critical tyrosine residues regulate the enzymatic and biological activity of Raf-1 kinase , 1993, Molecular and cellular biology.

[46]  J. Sedivy,et al.  Raf-1 protein kinase activates the NF-kappa B transcription factor by dissociating the cytoplasmic NF-kappa B-I kappa B complex. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[47]  J. Russo,et al.  Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. , 1990, Cancer research.

[48]  J. L. Bos,et al.  ras oncogenes in human cancer: a review. , 1989, Cancer research.

[49]  G. Todaro,et al.  Growth factors from murine sarcoma virus-transformed cells. , 1978, Proceedings of the National Academy of Sciences of the United States of America.