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Id3 is a novel regulator of p27kip1 mRNA in early G1 phase and is required for cell-cycle progression

Oncogene volume 26, pages 5772–5783 (2007)Cite this article

Abstract

P27kip is a key inhibitory protein of the cell-cycle progression, which is rapidly downregulated in early G1 phase by a post-translational mechanism involving the proteosomal degradation. In this study, using a wounding model that induces cell-cycle entry of human dermal fibroblasts, we demonstrate that p27mRNA is downregulated when cells progress into the G1 phase, and then it returns to its basal level when cells approach the S phase. By using a quantitative polymerase chain reaction screening we identified inhibitors of differentiation (Id3), a bHLH transcriptional repressor, as a candidate mediator accounting for p27 mRNA decrease. Id3 silencing, using an small interfering RNA approach, reversed the injury mediated p27 downregulation demonstrating that Id3 is involved in the transcriptional repression of p27. Reporter gene experiments and a chromatin immunoprecipitation assay showed that Id3 likely exerts its repressive action through ELK1 inhibition. By inhibiting early p27 downregulation, Id3 depletion blocked (i) the G1-phase progression as assessed by the inhibition of pRb phosphorylation and p130 degradation and (ii) the G1/S transition as observed by the inhibition of cyclin A induction, demonstrating that p27 mRNA decrease is required for cell proliferation. Apart from its effect on the early p27 diminution, Id3 appears also involved in the control of the steady-state level of p27 at the G1/S boundary. In conclusion, this study identifies a novel mechanism of p27 regulation which besides p27 protein degradation also implicates a transcriptional mechanism mediated by Id3.

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Acknowledgements

We are grateful to Drs JD Norton for generous gifts of the expression vectors encoding wt-Id3 and Id3-ala5, to Dr T Sakai for the plasmid containing p27 promoter, to Dr R Stein for the E12 and E47 constructs, to Dr C Sardet and JM Blanchard for the p27-null MEFs and to Dr R Hipskind for kindly providing the elk1 constructs. AA Chassot is recipient of a fellowship from the INSERM/Région PACA and from la Ligue Nationale contre le Cancer (LNCC). L Turchi is recipient of a fellowship of the Centre Hospitalier Universitaire (CHU) de Nice. This work was supported by grants of the Institut National de la Santé et la Recherche Médicale (INSERM), L'association pour la Recherche contre le Cancer, (ARC grant N° 5965 and 4500) and of the ‘Ministère de l'Education Nationale et de la Recherche’ (MENR) (ACI ACI: TS/0220045).

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Author notes

  1. A-A Chassot and L Turchi: These two authors equally contributed to this work.

Authors and Affiliations

  1. INSERM U634; Faculté de Médecine, Université Nice Sophia Antipolis, Nice cedex, France

    A-A Chassot, L Turchi, T Virolle, G Fitsialos, G Meneguzzi, R Buscà & G Ponzio

  2. INSERM U576, Hôpital de L'Archet, Université Nice Sophia Antipolis, Nice, France

    M Batoz & M Deckert

  3. CRBM-CNRS FRE 2593,, Montpellier, France

    V Dulic

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  1. A-A Chassot
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Correspondence to G Ponzio.

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Chassot, AA., Turchi, L., Virolle, T. et al. Id3 is a novel regulator of p27kip1 mRNA in early G1 phase and is required for cell-cycle progression. Oncogene 26, 5772–5783 (2007). https://doi.org/10.1038/sj.onc.1210386

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