Conserved sequences in the final intron of MDM2 are essential for the regulation of alternative splicing of MDM2 in response to stress. Exp Cell Res 2009 Nov 15;315(19):3419-32
Date
07/28/2009Pubmed ID
19631207DOI
10.1016/j.yexcr.2009.07.017Scopus ID
2-s2.0-70350405712 (requires institutional sign-in at Scopus site) 17 CitationsAbstract
Alternative splicing plays a fundamental role in generating proteome diversity and is critical in regulation of eukaryotic gene expression. It is estimated that 50% of disease-causing mutations alter splicing efficiency and/or patterns of splicing. An alternatively spliced form of murine double-minute 2, MDM2-ALT1, is associated with pediatric rhabdomyosarcoma (RMS) at high frequency in primary human tumors and RMS cell lines. We have identified that this isoform can be induced in response to specific types of stress (UV and cisplatin). However, the mechanism of alternative splicing of MDM2 in human cancer is unknown. Using UV and cisplatin to model alternative splicing of the MDM2 gene, we have developed a damage-inducible in vitro splicing system. This system employs an MDM2 minigene that mimics the damage-induced alternative splicing observed in vivo. Using this in vitro splicing system, we have shown that conserved intronic sequences in intron 11 of MDM2 are required for normal splicing. Furthermore, we showed that these intronic elements are also required for the regulated damage-induced alternative splicing of MDM2. The use of this novel damage-inducible system will allow for the systematic identification of regulatory elements and factors involved in the splicing regulation of the MDM2 gene in response to stress. This study has implications for identification of novel intervention points for development of future therapeutics for rhabdomyosarcoma.
Author List
Singh RK, Tapia-Santos A, Bebee TW, Chandler DSMESH terms used to index this publication - Major topics in bold
Alternative SplicingAnimals
Cisplatin
Conserved Sequence
Introns
Mice
Proto-Oncogene Proteins c-mdm2
Stress, Physiological
Ultraviolet Rays