Protein chemical shift assignments of the unbound and RNA-bound forms of the alternative splicing factor SUP-12 from C. elegans
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The splicing factor SUP-12 from Caenorhabditis elegans binds to regulatory RNA elements in pre-mRNA in order to generate tissue-specific alternative splicing for genes such as the fibroblast growth factor receptor egl-15. In nematode muscle cells, SUP-12 promotes the use of a mutually exclusive exon to impart variant binding specificity to the EGL-15 extracellular protein domain. Here we report the side chain and backbone 1H, 13C and 15N chemical shift assignments for the bacterially expressed RNA recognition motif domain from SUP-12, both in isolation as well as bound to a short RNA derived from the intron sequence between exon 4 and exon 5B of egl-15. Comparison of protein chemical shift values for both the backbone and side chain nuclei, coupled with secondary chemical shift analysis, reveal initial details of the RNA recognition.
KeywordsAlternative splicing Nematode Muscle RRM RNA
We thank Edward Sparks for assistance with cloning, as well as the Structural Biology Platform and the Analytical and Preparative Techniques Platform at the Institut Européen de Chimie et Biologie for additional assistance. This research was supported by funds from the Institut de Chimie des Substances Naturelles, the Aquitaine regional government, the Institut Européen de Chimie et Biologie, and the French national equipment network for NMR spectroscopy (TGE RMN THC Fr3050).