p300 is required for MyoD-dependent cell cycle arrest and muscle-specific gene transcription.
The nuclear phosphoprotein p300 is a new member of a family of 'co-activators' (which also includes the CREB binding protein CBP), that directly modulate transcription by interacting with components of the basal transcriptional machinery. Both p300 and CBP are targeted by the adenovirus E1A protein, and binding to p300 is required for E1A to inhibit terminal differentiation in both keratinocytes and myoblasts. Here we demonstrate that, in differentiating skeletal muscle cells, p300 physically interacts with the myogenic basic helix-loop-helix (bHLH) regulatory protein MyoD at its DNA binding sites. During muscle differentiation, MyoD plays a dual role: besides activating muscle-specific transcription, it induces permanent cell cycle arrest by up-regulating the cyclin-dependent kinase inhibitor p21. We show that p300 is involved in both these activities. Indeed, E1A mutants lacking the ability to bind p300 are greatly impaired in the repression of E-box-driven transcription, and p300 overexpression rescues the wild-type E1A-mediated repression. Moreover, p300 potentiates MyoD- and myogenin-dependent activation of transcription from E-box-containing reporter genes. We also provide evidence, obtained by microinjection of anti-p300 antibodies, that p300 is required for MyoD-dependent cell cycle arrest in either myogenic cells induced to differentiate or in MyoD-converted C3H10T1/2 fibroblasts, but is dispensable for maintenance of the postmitotic state of myotubes.
Puri
P L
PL
Fondazione Andrea Cesalpino, Università degli Studi di Roma La Sapienza, Rome, Italy.
Avantaggiati
M L
ML
Balsano
C
C
Sang
N
N
Graessmann
A
A
Giordano
A
A
Levrero
M
M
D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D002454:Cell Differentiation; D004247:DNA / Q000378:metabolism; D050881:E1A-Associated p300 Protein; D018507:Gene Expression Regulation, Developmental; D018257:Helix-Loop-Helix Motifs; D051379:Mice; D008845:Microinjections; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000254:growth & development*; D017570:MyoD Protein / Q000502:physiology*; D018008:Myogenin / Q000378:metabolism; D009687:Nuclear Proteins / Q000502:physiology*; D015534:Trans-Activators; D014157:Transcription Factors / Q000502:physiology*
D017570:MyoD Protein; C503801:Myog protein, mouse; D018008:Myogenin; D009687:Nuclear Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D004247:DNA; D050881:E1A-Associated p300 Protein; C494361:Ep300 protein, mouse