During the course of a large scale interference RNA (RNAi) screen in Caenorhabditis elegans, rsr-2 was identified as a genetic interactor of lin-35 Rb, the homolog of human Retinoblastoma. The rsr-2 gene encodes the ortholog of the human spliceosomal protein SRm300/SRRM2. In contrast to its yeast ortholog Cwc21, rsr-2 is essential for viability. Since strong inactivation of rsr-2 produces severe phenotypes, we took advantage of the mild effect of RNAi by feeding to study functions of rsr-2 during development. rsr-2 RNAi assays and genetic epistasis analysis locate rsr-2 within the germ line sex determination pathway but tiling arrays of rsr-2(RNAi) animals do not disclose significant splicing defects. By inmunofluorescence, we observe that an antibody specific for RSR-2 co-localizes with chromatin in germ line nuclei. Interestingly, ChIP-Seq experiments reveal that RSR-2 co-precipitates chromatin in a pattern similar to that of RNA Polymerase II. These ChIP-Seq assays also evidenced a splicing-independent recruitment of RSR-2 to chromatin and suggest that RSR-2 could have a role in regulating transcription. Moreover, we have explored the transcriptomes of rsr-2(RNAi) and prp-8(RNAi) L3 worms by RNA-Seq, which classified rsr-2 as a non-essential splicing factor. Altogether, our study shows that RSR-2 is a multifunctional protein that regulates development by influencing, and probably coupling, splicing and transcription.
|Date of Award||13 Jul 2012|
|Supervisor||Antoni Barbadilla Prados (Director) & Jose Joaquín Cerón (Director)|