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Nucleic Acids Research 2005 33(7):2332-2342; doi:10.1093/nar/gki530
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Published online 22 April 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

High frequency trans-splicing in a cell line producing spliced and polyadenylated RNA polymerase I transcripts from an rDNA-myc chimeric gene

Célia Chen, Nicole Fossar, Dominique Weil2, Marine Guillaud-Bataille1, Gisèle Danglot1, Brigitte Raynal, François Dautry2, Alain Bernheim1 and Olivier Brison*

Laboratoire de Génétique Oncologique, UMR 8125, Institut Gustave Roussy Villejuif, France 1Laboratoire de Génomique Cellulaire des Cancers, UMR 8125, Institut Gustave Roussy Villejuif, France 2Génétique Moléculaire des Fonctions Cellulaires, UPR 1983, Institut André Lwoff Villejuif, France

*To whom correspondence should be addressed at UMR 7147, Institut Curie, 26 rue d'Ulm, 75248 Paris Cedex 05, France. Tel: +33 1 42 34 66 68; Fax: +33 1 42 34 66 74; Email: olivier.brison{at}curie.fr

Received October 10, 2004. Revised March 15, 2005. Accepted April 4, 2005.

The 2G1MycP2Tu1 cell line was obtained following transfection of human colon carcinoma cells from the SW613-S cell line with a plasmid carrying a genomic copy of the human MYC gene. 2G1MycP2Tu1 cells produce MYC mRNAs and proteins of abnormal size. In order to analyze the structure of these abnormal products, a cDNA library constructed using RNA isolated from these cells was screened with a MYC probe. Fifty clones were studied by DNA sequencing. The results indicated that a truncated copy of the MYC gene had integrated into an rDNA transcription unit in 2G1MycP2Tu1 cells. This was confirmed by northern blot analysis, PCR amplification on genomic DNA and fluorescent in situ hybridization (FISH) experiments on metaphase chromosomes. 2G1MycP2Tu1 cells produce hybrid rRNA-MYC RNA molecules that are polyadenylated and processed by splicing reactions involving natural and cryptic splice sites. These transcripts are synthesized by RNA polymerase I, as confirmed by actinomycin D sensitivity experiments, suggesting that 3' end processing and splicing are uncoupled from transcription in this case. 2G1MycP2Tu1 cells also produce another type of chimeric mRNAs consisting of correctly spliced exons 2 and 3 of the MYC gene fused to one or more extraneous 5' exons by proper splicing to the acceptor sites of MYC exon 2. These foreign exons belong to 33 different genes, which are located on 14 different chromosomes. These observations and the results of FISH and Southern blotting experiments lead us to conclude that trans-splicing events occur at high frequency in 2G1MycP2Tu1 cells.

DDBJ/EMBL/GenBank accession nos AJ749659–AJ749668


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