The CBP co-activator stimulates E2F1/DP1 activity.
The cell cycle-regulating transcription factors E2F1/DP1 activate genes whose products are required for S phase progression. During most of the G1 phase, E2F1/DP1 activity is repressed by the retinoblastoma gene product RB, which directly contacts the E2F1 activation domain and silences it. The E2F1 activation domain has sequence similarity to the N-terminal activation domain of E1A(12S), which contains binding sites for CBP as well as RB. Here, we present evidence that the CBP protein directly contacts E2F1/DP1 and stimulates its activation capacity. We show that CBP interacts with the activation domain of E2F1 both in vitro and in vivo. Deletion of four residues from the E2F1 activation domain reduces CBP binding as well as transcriptional activation, but still allows the binding of RB and MDM2. This deletion removes residues which are conserved in the N-terminal activation domain of E1A and which are required for the binding of CBP to E1A. When the E1A N-terminus is used as a competitor in squelshing experiments it abolishes CBP-induced activation of E2F1/DP1, whereas an E1A mutant lacking CBP binding ability fails to do so. These results indicate that CBP can act as a coactivator for E2F1 and suggest that CBP recognises a similar motif within the E1A and E2F1 activation domains. The convergence of the RB and CBP pathways on the regulation of E2F1 activity may explain the cooperativity displayed by these proteins in mediating the biological functions of E1A. We propose a model in which E1A activates E2F not only by removing the RB repression but also by providing the CBP co-activator.
Trouche
D
D
Wellcome/CRC Institute and Department of Pathology, University of Cambridge, UK.
Cook
A
A
Kouzarides
T
T
D000595:Amino Acid Sequence; D001665:Binding Sites; D050882:CREB-Binding Protein; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D056465:Retinoblastoma-Binding Protein 1; D015534:Trans-Activators; D014157:Transcription Factors / Q000378:metabolism*
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D009687:Nuclear Proteins; D056465:Retinoblastoma-Binding Protein 1; D015534:Trans-Activators; D014157:Transcription Factors; D050882:CREB-Binding Protein