Manipulation of nonsense mediated decay identifies gene mutations in colon cancer Cells with microsatellite instability View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2004-01-22

AUTHORS

Yurij Ionov, Norma Nowak, Manuel Perucho, Sanford Markowitz, John K Cowell

ABSTRACT

Cancer cells showing microsatellite instability (MSI) demonstrate a high frequency of acquired frameshift mutations that result in the generation of nonsense mutations. RNA transcripts carrying these nonsense mutations are usually targeted for degradation through the nonsense mediated decay (NMD) pathway. Blocking this pathway with drugs such as emitine, results in the ‘stabilization’ of these mutant transcripts, which can now be detected on cDNA arrays. Unfortunately, emetine also induces a stress response that results in upregulation of additional transcripts which contribute to the analysis of the array. As a result, identifying which genes truly carry nonsense mutations is made more difficult. To overcome this, we have combined the emetine treatment with actinomycin D, which effectively prevents the upregulation of stress response genes while still stabilizing mutant transcripts. When we applied this modified approach to the analysis of MSI-positive colon cancer cells, we identified mutations in the UVRAG and p300 genes. More... »

PAGES

639-645

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/sj.onc.1207178

DOI

http://dx.doi.org/10.1038/sj.onc.1207178

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1044734677

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/14737099


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