Abstract
The oncogenic fusion protein p210 Bcr-Abl is causally associated with virtually all cases of chronic myelogenous leukemia. The wild-type Bcr product has several recognizable structural and functional motifs including a domain that contains guanine nucleotide exchange activity for Rho family GTPases (DH/PH domain). Although this domain is retained within p210 Bcr-Abl, it has no known signaling activities in vivo. Here we report that a fragment of Bcr that encodes the isolated DH/PH domain is a potent activator of the NF-kappaB transcription factor. Within the context of full length Bcr, this activity is regulated by proximal flanking sequences that suppress the DH/PH domain encoded guanine nucleotide exchange activity. NF-kappaB activation by Bcr is not mediated by nuclear translocation, but rather by p38 mitogen-activated protein kinase (MAPK)-dependent modification of the RelA/p65 transactivation domain. Although we were able to demonstrate that Bcr can function as an exchange factor for Cdc42 in vivo, NF-kappaB activation appears to occur via a Cdc42-independent mechanism. These studies constitute direct evidence that the Bcr RhoGEF domain can function in vivo, and identify a new signaling activity that may contribute to the transforming potential of p210 Bcr-Abl.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3T3 Cells
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Animals
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Binding Sites
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COS Cells
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Cell Nucleus / metabolism
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Electrophoretic Mobility Shift Assay
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Fusion Proteins, bcr-abl / chemistry
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Fusion Proteins, bcr-abl / metabolism
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Guanine Nucleotide Exchange Factors / metabolism*
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MAP Kinase Signaling System
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Mice
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Mitogen-Activated Protein Kinases / metabolism*
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NF-kappa B / antagonists & inhibitors
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NF-kappa B / chemistry
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NF-kappa B / metabolism*
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Oncogene Proteins / chemistry*
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Oncogene Proteins / metabolism*
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Promoter Regions, Genetic / genetics
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Protein Structure, Tertiary
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Protein-Tyrosine Kinases*
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Proto-Oncogene Proteins c-bcr
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Proto-Oncogene Proteins*
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Response Elements / genetics
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Rho Guanine Nucleotide Exchange Factors
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Transcriptional Activation
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Transfection
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Tumor Necrosis Factor-alpha / pharmacology
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cdc42 GTP-Binding Protein / metabolism
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p38 Mitogen-Activated Protein Kinases
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rac1 GTP-Binding Protein / metabolism
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rhoA GTP-Binding Protein / metabolism
Substances
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Guanine Nucleotide Exchange Factors
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NF-kappa B
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Oncogene Proteins
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Proto-Oncogene Proteins
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Rho Guanine Nucleotide Exchange Factors
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Tumor Necrosis Factor-alpha
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Protein-Tyrosine Kinases
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Fusion Proteins, bcr-abl
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Bcr protein, mouse
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Proto-Oncogene Proteins c-bcr
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Mitogen-Activated Protein Kinases
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p38 Mitogen-Activated Protein Kinases
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cdc42 GTP-Binding Protein
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rac1 GTP-Binding Protein
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rhoA GTP-Binding Protein