Abstract
The small cell lung carcinoma (SCLC) cell line DMS-79 has been used as a model for studying the molecular mechanism underlying the ectopic ACTH syndrome. We previously showed that two domains of the human Proopiomelanocortin (POMC) gene promoter were specifically active in DMS-79 cells. The present study focuses on the more distal one, Domain IV (−376/−417). DNaseI footprinting experiments identified a single binding site for DMS-79 cell proteins in this domain. Gel-shift and sequence analysis indicated that E2F proteins might bind this site. Indeed, proteins from DMS-79 cells which bind this site (i) have in vitro DNA binding properties indistinguishable from those of E2F proteins (ii) form, like E2F proteins, multiprotein complexes which can be dissociated by sodium deoxycholate and (iii) are recognized by antibodies directed against E2F proteins. Further, we show that the rat POMC distal promoter domain contains a homologous sequence which constitutes a natural mutant of the human POMC E2F binding site, since it does not bind E2F. We show by transient transfection that this natural mutant, in the context of the rat POMC promoter, is not active in DMS-79 cells by contrast to the human POMC E2F binding site. We conclude that E2F binding is required for the activity of Domain IV in DMS-79 cells and contributes to the expression of the POMC gene in SCLC. Further studies are required to elucidate the role of E2F factors in POMC gene transcription in SCLC cells, but our results have identified mechanisms different from those in pituitary corticotroph cells that are used by these SCLC tumor cells.
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Acknowledgements
We are indebted to Dr GD Sorenson and Dr O Pettengill for having provided us with the DMS-79 cell line. We wish to thank Drs DN Orth for his valuable comments on the manuscript and M Raymondjean for helpful discussions and comments. We thank Dr J Drouin for rat POMC plasmids and Drs G Brun and L Loiseau for chDP-1 and chE2F-1 plasmids. This work was supported in part by INSERM CJF92-08 and the Association pour la Recherche Contre le Cancer, (contract 6501 to YdK). A Picon is the recipient of a doctoral fellowship of the Association pour la Recherche Contre le Cancer.
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Picon, A., Bertagna, X. & de Keyzer, Y. Analysis of the human proopiomelanocortin gene promoter in a small cell lung carcinoma cell line reveals an unusual role for E2F transcription factors. Oncogene 18, 2627–2633 (1999). https://doi.org/10.1038/sj.onc.1202635
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DOI: https://doi.org/10.1038/sj.onc.1202635
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