Abstract
The protein product of the retinoblastoma tumor suppressor gene (RE) has been demonstrated to bind c-Myc protein (Myc) in vitro. To determine whether RE regulates Myc transcriptional activity in vivo, GAL4-Myc chimeric expression plasmids were generated and cotransfected with a RE expression plasmid and a GAL4-dependent reporter plasmid. RE stimulated GAL4-Myc-mediated transcription, dependent upon a domain(s) in the amino-terminus of Myc. The stimula tion of Myc-mediated transcription by RE was cell-type specific and was inhibited by SV40 T-antigen, but not by a T-antigen mutant defective in RE-binding. Moreover, RE mutants containing mutations in domain E of RE pocket were significantly reduced in their ability to stimulate GAL4-Myc mediated transcription. To determine whether RE and Myc interact in vivo either directly or indirectly, a two hybrid system was used where GAL4-Rb and Myc-VP16 expression constructs were cotransfected with a GAL4-dependent reporter plasmid. A significant increase of GAL4-dependent transcription was observed, dependent upon the presence of both GAL4-Rb and Myc-VP16 fusion proteins. Mutational analysis of the Myc-VP16 chimeric proteins suggests that the amino-terminus of Myc is essential for the interaction with RE. These results demonstrate that RE can regulate Myc-mediated transcription in vivo in a cell-type specific manner through protein-protein interactions.
Original language | English (US) |
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Pages (from-to) | 381-387 |
Number of pages | 7 |
Journal | Oncogene |
Volume | 10 |
Issue number | 2 |
State | Published - Jan 19 1995 |
Externally published | Yes |
Keywords
- Myc
- Retinoblastoma protein
- Transcription