Repression of c-Myc and inhibition of G1 exit in cells conditionally overexpressing p300 that is not dependent on its histone acetyltransferase activity.
p300 and cAMP response element binding protein (CREB)-binding protein (CBP) are two highly homologous, conserved transcriptional coactivators, and histone acetyltransferases (HATs) that link chromatin remodeling with transcription. Cell transformation by viral oncogene products such as adenovirus E1A and SV40 large T antigen depends on their ability to inactivate p300 and CBP. To investigate the role of p300 in cell-cycle progression, we constructed stable rat cell lines, which conditionally overexpress p300 from a tetracycline-responsive promoter. When p300 was induced in these cells, serum-stimulated S-phase entry was significantly inhibited. The inhibition of S-phase induction was associated with down-regulation of c-Myc, but not of c-Fos or c-Jun. Simultaneous overexpression of c-Myc and p300 before serum stimulation reversed the inhibition of S-phase induction to a significant level, indicating that the inhibition of c-Myc to a large extent is responsible for the p300 inhibition of G1 exit. Similar studies with stable rat cell lines that overexpress a mutant p300, which lacks the HAT activity, showed that the intrinsic HAT activity of p300 is not required for the negative regulation of c-Myc or G1. These findings, and our previously published results (Kolli, S., Buchmann, A. M., Williams, J., Weitzman, S. & Thimmapaya, B. (2001) Proc. Natl. Acad. Sci. USA 98, 4646-4651), establish an important negative regulatory role for p300 in c-Myc expression that may be important in maintaining the cells in the G0/G1 phase of the cell cycle.
Baluchamy
Sudhakar
S
Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
Rajabi
Hasan N
HN
Thimmapaya
Rama
R
Navaraj
Arunasalam
A
Thimmapaya
Bayar
B
D000123:Acetyltransferases / Q000378:metabolism*; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D002453:Cell Cycle; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D015536:Down-Regulation; D050881:E1A-Associated p300 Protein; D016193:G1 Phase; D005822:Genetic Vectors; D006657:Histones / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D011233:Precipitin Tests; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D051381:Rats; D016196:S Phase; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism*; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured
D006657:Histones; D009687:Nuclear Proteins; D016271:Proto-Oncogene Proteins c-myc; D015534:Trans-Activators; D004247:DNA; D000123:Acetyltransferases; D050881:E1A-Associated p300 Protein; C494362:Ep300 protein, rat