Damage-inducible intragenic demethylation of the human TP53 tumor suppressor gene is associated with transcription from an alternative intronic promoter
Wild-type TP53 exons 5-8 contain CpG dinucleotides that are prone to methylation-dependent mutation during carcinogenesis, but the regulatory effects of methylation affecting these CpG sites are unclear. To clarify this, we first assessed site-specific TP53 CpG methylation in normal and transformed cells. Both DNA damage and cell ageing were associated with site-specific CpG demethylation in exon 5 accompanied by induction of a truncated TP53 isoform regulated by an adjacent intronic promoter (P2). We then synthesized novel synonymous TP53 alleles with divergent CpG content but stable encodement of the wild-type polypeptide. Expression of CpG-enriched TP53 constructs selectively reduced production of the full-length transcript (P1), consistent with a causal relationship between intragenic demethylation and transcription. 450K methylation comparison of normal (TP53-wildtype) and cancerous (TP53-mutant) human cells and tissues revealed focal cancer-associated declines in CpG methylation near the P1 transcription start site, accompanied by rises near the alternate exon 5 start site. These data confirm that site-specific changes of intragenic TP53 CpG methylation are extrinsically inducible, and suggest that human cancer progression is mediated in part by dysregulation of damage-inducible intragenic CpG demethylation that alters TP53 P1/P2 isoform expression. (c) 2015 The Authors. Molecular Carcinogenesis Published by Wiley Periodicals, Inc.
|ISBN||1098-2744 (Electronic) 0899-1987 (Linking)|
|Authors||Blackburn, J.; Roden, D. L.; Ng, R.; Wu, J.; Bosman, A.; Epstein, R. J.;|
|Publisher Name||MOLECULAR CARCINOGENESIS|
|URL link to publisher's version||http://www.ncbi.nlm.nih.gov/pubmed/26676339|
|OpenAccess link to author's accepted manuscript version||https://publications.gimr.garvan.org.au/open-access/13440|