Heterochromatin silencing of p53 target genes by a small viral protein

Abstract

The transcription factor p53 (also known as TP53) guards against tumour and virus replication and is inactivated in almost all cancers. p53-activated transcription of target genes is thought to be synonymous with the stabilization of p53 in response to oncogenes and DNA damage. During adenovirus replication, the degradation of p53 by E1B-55k is considered essential for p53 inactivation, and is the basis for p53-selective viral cancer therapies. Here we reveal a dominant epigenetic mechanism that silences p53-activated transcription, irrespective of p53 phosphorylation and stabilization. We show that another adenoviral protein, E4-ORF3, inactivates p53 independently of E1B-55k by forming a nuclear structure that induces de novo H3K9me3 heterochromatin formation at p53 target promoters, preventing p53–DNA binding. This suppressive nuclear web is highly selective in silencing p53 promoters and operates in the backdrop of global transcriptional changes that drive oncogenic replication. These findings are important for understanding how high levels of wild-type p53 might also be inactivated in cancer as well as the mechanisms that induce aberrant epigenetic silencing of tumour-suppressor loci. Our study changes the longstanding definition of how p53 is inactivated in adenovirus infection and provides key insights that could enable the development of true p53-selective oncolytic viral therapies.

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Figure 1: p53 is induced and phosphorylated in ΔE1B-55k infection but p53 activity is dominantly suppressed.
Figure 2: E4-ORF3 inactivates p53 independently of E1B-55k and p53 degradation.
Figure 3: E4-ORF3 induces heterochromatin formation and prevents p53–DNA binding at endogenous promoters.
Figure 4: E4-ORF3 forms a nuclear scaffold that specifies heterochromatin assembly and H3K9 trimethylation at p53 target promoters.
Figure 5: p53 transcriptional targets are silenced selectively in the backdrop of global transcriptional changes that drive oncogenic cellular and viral replication.

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Acknowledgements

We thank J. Fitzpatrick and the Waitt Advanced Biophotonics Center for assistance with imaging and analysis, J. Karlseder, I. Verma, T. Hunter, R. Shaw and the O’Shea laboratory for critical reading of this manuscript, L. Haro, S. Panda, R. O’Sullivan and A. Rodriguez for advice and protocols, and P. Branton and D. Ornelles for viruses. C.C.O. acknowledges funding from the Alliance of Cancer Gene Therapy, the American Cancer Society, the Sontag Foundation and the Beckman Foundation. This work was supported by R01CA137094 from the National Cancer Institute.

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C.S. performed the p53 activation and virus studies, including immunoblotting, RT–qPCR and microarray experiments. F.E.E. performed all chromatin immunoprecipitation and immunofluorescence studies. K.C.E. performed the luciferase assays, E4-ORF3 sufficiency and complementation, and assisted C.S. with viral mutant studies. C.C.O. analysed the array data and wrote the paper with contributions from all authors. C.C.O. was responsible for the overall conceptual design and supervision of the studies.

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Correspondence to Clodagh C. O’Shea.

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The authors declare no competing financial interests.

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Microarray data are deposited in NCBI's Gene Expression Omnibus (GSE20607).

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Soria, C., Estermann, F., Espantman, K. et al. Heterochromatin silencing of p53 target genes by a small viral protein. Nature 466, 1076–1081 (2010). https://doi.org/10.1038/nature09307

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