Transgenic alternative-splicing reporters reveal tissue-specific expression profiles and regulation mechanisms in vivo.
Alternative splicing of pre-mRNAs allows multicellular organisms to create a huge diversity of proteomes from a finite number of genes. But extensive studies in vitro or in cultured cells have not fully explained the regulation mechanisms of tissue-specific or developmentally regulated alternative splicing in living organisms. Here we report a transgenic reporter system that allows visualization of expression profiles of mutually exclusive exons in Caenorhabditis elegans. Reporters for egl-15 exons 5A and 5B showed tissue-specific profiles, and we isolated mutants defective in the tissue specificity. We identified alternative-splicing defective-1 (asd-1), encoding a new RNA-binding protein of the evolutionarily conserved Fox-1 family, as a regulator of the egl-15 reporter. Furthermore, an asd-1;fox-1 double mutant was defective in the expression of endogenous egl-15 (5A) and phenocopied egl-15 (5A) mutant. This transgenic reporter system can be a powerful experimental tool for the comprehensive study of expression profiles and regulation mechanisms of alternative splicing in metazoans.
Kuroyanagi
Hidehito
H
School of Biomedical Science, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. kuroyana.end@tmd.ac.jp
Kobayashi
Tetsuo
T
Mitani
Shohei
S
Hagiwara
Masatoshi
M
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D020869:Gene Expression Profiling; D017930:Genes, Reporter / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics*
C515945:Asd-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; C112493:EGL-15 protein, C elegans; C117552:FOX-1 protein, C elegans; D016601:RNA-Binding Proteins; D017468:Receptors, Fibroblast Growth Factor