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The CDKN 2A/p16 INK 4a 5′ UTR sequence and translational regulation: impact of novel variants predisposing to melanoma
Author(s) -
Andreotti Virginia,
Bisio Alessandra,
Bressacde Paillerets Brigitte,
Harland Mark,
Cabaret Odile,
NewtonBishop Julia,
Pastorino Lorenza,
Bruno William,
Bertorelli Roberto,
De Sanctis Veronica,
Provenzani Alessandro,
Menin Chiara,
Fronza Gilberto,
Queirolo Paola,
Spitale Robert C.,
BianchiScarrà Giovanna,
Inga Alberto,
Ghiorzo Paola
Publication year - 2016
Publication title -
pigment cell and melanoma research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 105
eISSN - 1755-148X
pISSN - 1755-1471
DOI - 10.1111/pcmr.12444
Subject(s) - untranslated region , biology , microbiology and biotechnology , messenger rna , polyadenylation , translation (biology) , five prime untranslated region , internal ribosome entry site , genetics , gene
Summary Many variants of uncertain functional significance in cancer susceptibility genes lie in regulatory regions, and clarifying their association with disease risk poses significant challenges. We studied 17 germline variants (nine of which were novel) in the CDKN 2A 5′ UTR with independent approaches, which included mono and bicistronic reporter assays, Western blot of endogenous protein, and allelic representation after polysomal profiling to investigate their impact on CDKN 2A mRNA translation regulation. Two of the novel variants (c.‐27del23, c.‐93‐91del AGG ) were classified as causal mutations (score ≥3), along with the c.‐21C>T, c.‐34G>T, and c.‐56G>T, which had already been studied by a subset of assays. The novel c.‐42T>A as well as the previously described c.‐67G>C were classified as potential mutations (score 1 or 2). The remaining variants (c.‐14C>T, c.‐20A>G, c.‐25C>T+c.‐180G>A, c.‐30G>A, c.‐40C>T, c.‐45G>A, c.‐59C>G, c.‐87T>A, c.‐252A>T) were classified as neutral (score 0). In conclusion, we found evidence that nearly half of the variants found in this region had a negative impact on CDKN 2A mRNA translation, supporting the hypothesis that 5′ UTR can act as a cellular Internal Ribosome Entry Site ( IRES ) to modulate p16 INK4a translation.