Abstract
CREB binding protein (CBP) plays a central role in cell differentiation and proliferation, interacting with a large number of nuclear factors. To find novel nuclear factors associating with CBP, we have carried out yeast two-hybrid screening of human chondrocyte cDNA library using the C/H3 region of CBP as a bait and cloned CDK4 binding protein p34SEI-1, the recently found cell cycle regulator. The association of p34SEI-1 with CBP was confirmed in vitro by GST pull-down assay and in vivo by coimmunoprecipitation. Results of the immunofluorescence assay also supported the association of p34SEI-1 and CBP. In reporter assay using CRE promoter, p34SEI-1 strongly suppressed CREB-mediated transcription, and this suppression was overcome by excess amount of CBP, but not by CBPDeltaCH3. It is suggested that the association of p34SEI-1 and CBP is not only involved in cell cycle regulation by CBP, but also have some effect on other CBP-dependent transcription.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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CREB-Binding Protein
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Cell Cycle
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Cell Line
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Chondrocytes / metabolism
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Cloning, Molecular
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Cyclic AMP Response Element-Binding Protein / genetics
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Cyclic AMP Response Element-Binding Protein / metabolism*
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DNA, Complementary / genetics
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Humans
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In Vitro Techniques
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Macromolecular Substances
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Nuclear Proteins / chemistry
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Trans-Activators / chemistry
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Trans-Activators / genetics
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Trans-Activators / metabolism*
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Transcription Factors
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Transcription, Genetic
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Two-Hybrid System Techniques
Substances
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Cyclic AMP Response Element-Binding Protein
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DNA, Complementary
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Macromolecular Substances
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Nuclear Proteins
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Peptide Fragments
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Recombinant Fusion Proteins
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SERTAD1 protein, human
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Trans-Activators
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Transcription Factors
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CREB-Binding Protein
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CREBBP protein, human