Independent regulation of P53 stabilisation and activation after Rb deletion in primary epithelial cells.
We have previously reported that deletion of the retinoblastoma gene Rb leads to rapid but transient p53 stabilisation. We investigated here the pathways involved. We show that upon Rb-deletion dysregulated E2F activates p19ARF expression that localises in the nucleoli. There it interacts with MDM2, leading to P53 stabilisation. At the same time, ATR is activated, activating CHK1 that may phosphorylate P53 but also contribute to inhibition of MnSOD expression leading to accumulation of ROS (reactive oxygen species) and subsequent DNA injury, which in turn maintains ATR/CHK1 activated. However, from 72 h after Rb deletion, NPM interacts with P19ARF and concomitantly the interaction between p19ARF and MDM2 decreases leading to a return to P53 degradation. This occurs despite the persistence of the DNA damage response pathways. We therefore observe in primary cells not subjected to exogenous gene expression or exogenous DNA damaging treatment, activation of 2 concomitant pathways of activation of P53 that are dealt with in independent manner: an oncogenic pathway with rapid activation of ARF which is 'switched off' downstream of p19ARF activation after 72 h of induction and a DNA damage response pathway keeping a low level of transcriptionally active P53 sufficient to deal with a physiological elevation of oxidative DNA injury. A possible connection between the two pathways is discussed.
Division of Pathology, Queen's Medical Research Institute, 49 Little France Crescent, Edinburgh, EH16 4TJ Scotland, UK. louise.treanor@stjude.org
Louise
L
Treanor
Christopher
C
Bellamy
David J
DJ
Harrison
Sandrine
S
Prost
D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism*; D017353:Gene Deletion; D022781:Hepatocytes / Q000378:metabolism / Q000502:physiology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D018384:Oxidative Stress / Q000235:genetics / Q000502:physiology; D055550:Protein Stability; D016160:Retinoblastoma Protein / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D013482:Superoxide Dismutase / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism* / Q000502:physiology*
C417743:Cdkn2a protein, mouse; D019941:Cyclin-Dependent Kinase Inhibitor p16; D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53; D013482:Superoxide Dismutase