Abstract
Levels of the transcription factor B-myb must be down-regulated to allow terminal differentiation of neuroectodermal cells and yet its constitutive expression induces early markers of neural differentiation. Thus, we investigated potential mechanisms of enhanced B-myb activity in early stages of neural differentiation. We report here that B-myb expression does not decrease, cyclin A and Sp1 levels remain constant while p21 levels increase continuously upon retinoic acid-induced differentiation of the LAN-5 neuroblastoma cell line. In contrast, cyclin D1 expression is down-regulated at the onset of the differentiative process by protein destabilization. Luciferase assays of promoter activity indicate that B-myb-dependent transactivation is enhanced in LAN-5 cells treated with retinoic acid (RA) for 24 h. The enhancement is independent from cyclin A but is suppressed by a degradation-resistant mutant form of cyclin D1. The importance of cyclin D1 in controlling B-myb activity is further suggested by co-immunoprecipitation experiments, showing that the amount of cyclin D1 co-immunoprecipitated with B-myb decreased after RA treatment. Thus, B-myb may play an active role in the early stages of differentiation when its transactivation activity is enhanced as a consequence of cyclin D1 down-modulation.
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Abbreviations
- DTT:
-
dithiothreitol
- GSK-3β:
-
glycogen synthase kinase 3 beta
- NB:
-
neuroblastoma
- RT–PCR:
-
reverse transcriptase- polymerase chain reaction
- SDS–PAGE:
-
polyacrylamide gel electrophoresis in presence of sodium dodecyl sulphate
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Acknowledgements
This work was partly supported by grants of the Associazione Italiana per la Ricerca sul Cancro (AIRC) and of the Associazione Italiana per la Lotta al Neuroblastoma (to G.Raschellà) and of the National Institute of Health (NIH) (to B Calabretta). V Cesi is recipient of a fellowship from the Fondazione Adriano Buzzati-Traverso. B Tanno is recipient of a fellowship from the Fondazione Italiana per la Ricerca sul Cancro (FIRC).
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Cesi, V., Tanno, B., Vitali, R. et al. Cyclin D1-dependent regulation of B-myb activity in early stages of neuroblastoma differentiation. Cell Death Differ 9, 1232–1239 (2002). https://doi.org/10.1038/sj.cdd.4401103
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DOI: https://doi.org/10.1038/sj.cdd.4401103
