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dc.contributor.authorWilliams, David Sen
dc.contributor.authorBird, Matthew Jen
dc.contributor.authorJorissen, Robert Nen
dc.contributor.authorYu, Yen Linen
dc.contributor.authorWalker, Franscesaen
dc.contributor.authorZhang, Hui Huaen
dc.contributor.authorNice, Edouard Cen
dc.contributor.authorBurgess, Antony Wen
dc.identifier.citationPlos One 2010; 5(12): e16012en
dc.description.abstractFrameshift mutations in microsatellite instability high (MSI-High) colorectal cancers are a potential source of targetable neo-antigens. Many nonsense transcripts are subject to rapid degradation due to nonsense-mediated decay (NMD), but nonsense transcripts with a cMS in the last exon or near the last exon-exon junction have intrinsic resistance to nonsense-mediated decay (NMD). NMD-resistant transcripts are therefore a likely source of expressed mutant proteins in MSI-High tumours.Using antibodies to the conserved N-termini of predicted mutant proteins, we analysed MSI-High colorectal cancer cell lines for examples of naturally expressed mutant proteins arising from frameshift mutations in coding microsatellites (cMS) by immunoprecipitation and Western Blot experiments. Detected mutant protein bands from NMD-resistant transcripts were further validated by gene-specific short-interfering RNA (siRNA) knockdown. A genome-wide search was performed to identify cMS-containing genes likely to generate NMD-resistant transcripts that could encode for antigenic expressed mutant proteins in MSI-High colon cancers. These genes were screened for cMS mutations in the MSI-High colon cancer cell lines.Mutant protein bands of expected molecular weight were detected in mutated MSI-High cell lines for NMD-resistant transcripts (CREBBP, EP300, TTK), but not NMD-sensitive transcripts (BAX, CASP5, MSH3). Expression of the mutant CREBBP and EP300 proteins was confirmed by siRNA knockdown. Five cMS-bearing genes identified from the genome-wide search and without existing mutation data (SFRS12IP1, MED8, ASXL1, FBXL3 and RGS12) were found to be mutated in at least 5 of 11 (45%) of the MSI-High cell lines tested.NMD-resistant transcripts can give rise to expressed mutant proteins in MSI-High colon cancer cells. If commonly expressed in primary MSI-High colon cancers, MSI-derived mutant proteins could be useful as cancer specific immunological targets in a vaccine targeting MSI-High colonic tumours.en
dc.subject.otherCell Line, Tumoren
dc.subject.otherCodon, Nonsenseen
dc.subject.otherColonic Neoplasms.geneticsen
dc.subject.otherComputational Biology.methodsen
dc.subject.otherDNA Repairen
dc.subject.otherEpitope Mappingen
dc.subject.otherFrameshift Mutationen
dc.subject.otherGene Expression Regulation, Neoplasticen
dc.subject.otherHLA-A Antigens.geneticsen
dc.subject.otherMicrosatellite Instabilityen
dc.subject.otherMicrosatellite Repeatsen
dc.subject.otherProtein Structure, Tertiaryen
dc.titleNonsense mediated decay resistant mutations are a source of expressed mutant proteins in colon cancer cell lines with microsatellite instability.en
dc.typeJournal Articleen
dc.identifier.journaltitlePlos Oneen
dc.identifier.affiliationEpithelial Biochemistry Laboratory, Ludwig Institute for Cancer Research, Melbourne Branch, Parkville, Victoria, Australiaen
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