The execution of the transcriptional axis mutant p53, E2F1 and ID4 promotes tumor neo-angiogenesis

Abstract

ID4 (inhibitor of DNA binding 4) is a member of a family of proteins that function as dominant-negative regulators of basic helix-loop-helix transcription factors. Growing evidence links ID proteins to cell proliferation, differentiation and tumorigenesis. Here we identify ID4 as a transcriptional target of gain-of-function p53 mutants R175H, R273H and R280K. Depletion of mutant p53 protein severely impairs ID4 expression in proliferating tumor cells. The protein complex mutant p53–E2F1 assembles on specific regions of the ID4 promoter and positively controls ID4 expression. The ID4 protein binds to and stabilizes mRNAs encoding pro-angiogenic factors IL8 and GRO-α. This results in the increase of the angiogenic potential of cancer cells expressing mutant p53. These findings highlight the transcriptional axis mutant p53, E2F1 and ID4 as a still undefined molecular mechanism contributing to tumor neo-angiogenesis.

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Figure 1: ID4 expression is transcriptionally induced by mutant p53.
Figure 2: Mutant p53 drives the expression of ID4 and associates with its promoter.
Figure 3: Transcriptionally competent protein complexes containing mutant p53 assemble in vivo on ID4 promoter.
Figure 4: E2F1 is required for mutant p53-mediated ID4 transcription.
Figure 5: Mutant p53 and ID4 expression promote angiogenesis.
Figure 6: ID4 promotes IL8 and GRO-α expression.
Figure 7: ID4 binds and stabilizes IL-8 and GRO-α mRNAs.
Figure 8: Expression of ID4 protein pairs with p53 positivity and high microvessel density (MVD) in breast cancer (BC) specimens.

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Acknowledgements

We are grateful to J.A. Kessler (Northwestern University's Feinberg School of Medicine) and L. Lania (University of Naples Federico II) for providing plasmids. We also thank G. Starace (Italian National Research Council (CNR)) for cell sorting. This work was supported by European Community (EC) FP6 “Active p53” and “Mutant p53” consortia. This publication reflects the authors' views and not necessarily those of the EC. The EC is not liable for any use that may be made of the information contained herein. Support by Associazione Italiana per la Ricerca sul Cancro (AIRC)-Rome Oncogenic Centre (ROC) to the oncogenomic platform, AIRC to G.B., S.S., F.F. and D.D.B., Lega Italiana Tumori to S.S., Ministero della Sanità and Alleanza contro il cancro is greatly appreciated. S.D.O. holds a fellowship from Italian Association for Cancer Research (FIRC).

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G.F. and S.D.O. performed microarrays, transcriptional and mRNA stability studies, chromatin and ribonucleoprotein immunoprecipitations; E.D. and T.S. performed bioinformatic analysis of microarray data; F.F. performed studies on polysomal cell fractions; M.M. and E.M. performed stainings on breast cancer specimens; I.T. and P.M. performed statistical analysis of breast cancer immunohistochemical data; D.D.B. and D.T. performed angiogenic assays. G.B., G.F. and S.S. supervised the project.

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Correspondence to Giovanni Blandino.

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Fontemaggi, G., Dell'Orso, S., Trisciuoglio, D. et al. The execution of the transcriptional axis mutant p53, E2F1 and ID4 promotes tumor neo-angiogenesis. Nat Struct Mol Biol 16, 1086–1093 (2009). https://doi.org/10.1038/nsmb.1669

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