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RSR-2, the Caenorhabditis elegans Ortholog of Human Spliceosomal Component SRm300/SRRM2, Regulates Development by Influencing the Transcriptional Machinery Fontrodona, Laura Porta de la Riva, Montserrat Morán, Tomás Niu, Wei Diaz, Mònica Aristizábal Corrales, David Villanueva Garatachea, Alberto Schwartz Navarro, Simó Reinke, Valerie Cerón Madrigal, Julián Expressió gènica Cromatina Gene expression Chromatin Protein components of the spliceosome are highly conserved in eukaryotes and can influence several steps of the gene expression process. RSR-2, the Caenorhabditis elegans ortholog of the human spliceosomal protein SRm300/SRRM2, is essential for viability, in contrast to the yeast ortholog Cwc21p. We took advantage of mutants and RNA interference (RNAi) to study rsr-2 functions in C. elegans, and through genetic epistasis analysis found that rsr-2 is within the germline sex determination pathway. Intriguingly, transcriptome analyses of rsr-2(RNAi) animals did not reveal appreciable splicing defects but instead a slight global decrease in transcript levels. We further investigated this effect in transcription and observed that RSR-2 colocalizes with DNA in germline nuclei and coprecipitates with chromatin, displaying a ChIP-Seq profile similar to that obtained for the RNA Polymerase II (RNAPII). Consistent with a novel transcription function we demonstrate that the recruitment of RSR-2 to chromatin is splicing-independent and that RSR-2 interacts with RNAPII and affects RNAPII phosphorylation states. Proteomic analyses identified proteins associated with RSR-2 that are involved in different gene expression steps, including RNA metabolism and transcription with PRP-8 and PRP-19 being the strongest interacting partners. PRP-8 is a core component of the spliceosome and PRP-19 is the core component of the PRP19 complex, which interacts with RNAPII and is necessary for full transcriptional activity. Taken together, our study proposes that RSR-2 is a multifunctional protein whose role in transcription influences C. elegans development. 2018-11-27T09:23:29Z 2018-11-27T09:23:29Z 2013-06-06 2018-07-24T12:48:17Z info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Reproducció del document publicat a: https://doi.org/10.1371/journal.pgen.1003543 PLoS Genetics, 2013, vol. 9, num. 6, p. e1003543 https://doi.org/10.1371/journal.pgen.1003543 info:eu-repo/grantAgreement/EC/FP7/206584/EU//CANCEROMICS cc by (c) Fontrodona et al., 2013 http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess Public Library of Science (PLoS) Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL))