Regulation of CREB-mediated transcription by association of CDK4 binding protein p34SEI-1 with CBP.
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.
Hirose
Takuji
T
Department of Genome Science, Institute of Medical Science, St. Marianna University School of Medicine, Kawasaki 216-8512, Japan.
Fujii
Ryouji
R
Nakamura
Hiroshi
H
Aratani
Satoko
S
Fujita
Hidetoshi
H
Nakazawa
Minako
M
Nakamura
Kohzo
K
Nishioka
Kusuki
K
Nakajima
Toshihiro
T
D050882:CREB-Binding Protein; D002453:Cell Cycle; D002460:Cell Line; D019902:Chondrocytes / Q000378:metabolism; D003001:Cloning, Molecular; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D017362:Cyclic AMP Response Element-Binding Protein; D018076:DNA, Complementary; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C402464:SERTAD1 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D050882:CREB-Binding Protein; C494363:CREBBP protein, human