High-Throughput Screening for Small Molecule Modulators of FGFR2-IIIb Pre-mRNA Splicing
Pre-mRNA splicing is the process in which a ribonucleoprotein complex called the spliceosome removes introns from a primary transcript and ligates its exons together . The spliced exons produce an mRNA containing the mature protein coding sequence. The splicing process is remarkably consistent in its choice of splice sites, and these choices can be regulated to allow different mRNA sequences to arise from the same pre-mRNA transcript, in a process called alternative splicing. In this way, genes often yield multiple mRNAs and encoded proteins to increase diversity of eukaryotic proteomes. Alternative splicing patterns often show developmental and tissue-specific expression. Misregulation of splicing is seen in human disease and can be a direct cause of pathology . Splicing regulatory mechanisms thus present a promising target for therapeutic intervention.
KeywordsSplice Factor Dual Fluorescence Reporter Cell Line Exon Inclusion Cell Signaling Cascade
- 17.Newman EA, Muh SJ, Hovhannisyan RH, Warzecha CC, Jones RB, McKeehan WL, Carstens RP (2006) Identification of RNA-binding proteins that regulate FGFR2 splicing through the use of sensitive and specific dual color fluorescence minigene assays. RNA 12(6):1129–1141. doi:10.1261/rna.34906CrossRefPubMedGoogle Scholar
- 19.Soret J, Bakkour N, Maire S, Durand S, Zekri L, Gabut M, Fic W, Divita G, Rivalle C, Dauzonne D, Nguyen CH, Jeanteur P, Tazi J (2005) Selective modification of alternative splicing by indole derivatives that target serine-arginine-rich protein splicing factors. Proc Natl Acad Sci USA 102(24):8764–8769. doi:10.1073/pnas.0409829102CrossRefPubMedGoogle Scholar
- 22.Kaida D, Motoyoshi H, Tashiro E, Nojima T, Hagiwara M, Ishigami K, Watanabe H, Kitahara T, Yoshida T, Nakajima H, Tani T, Horinouchi S, Yoshida M (2007) Spliceostatin A targets SF3b and inhibits both splicing and nuclear retention of pre-mRNA. Nat Chem Biol 3(9):576–583. doi:10.1038/nchembio.2007.18CrossRefPubMedGoogle Scholar
- 24.Boutz PL, Stoilov P, Li Q, Lin CH, Chawla G, Ostrow K, Shiue L, Ares M Jr, Black DL (2007) A post-transcriptional regulatory switch in polypyrimidine tract-binding proteins reprograms alternative splicing in developing neurons. Genes Dev 21(13):1636–1652. doi:10.1101/gad.1558107CrossRefPubMedGoogle Scholar