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Biochem Biophys Res Commun ; 497(2): 597-604, 2018 03 04.
Article in English | MEDLINE | ID: mdl-29454968

ABSTRACT

Transcription and pre-mRNA splicing are complex, coupled processes that involve transcriptional co-regulators. Ess2 (also termed Dgcr14) is a nuclear protein that enhances the transcriptional activity of retinoic acid receptor-related orphan receptor gamma/gamma-t (Rorγ/γt). Ess2 is also a component of the spliceosomal C complex (containing U2, U5 and U6 snRNAs). However, the domains in Ess2 that function in splicing and transcription have not been identified. To elucidate the roles of Ess2 in splicing and transcription, we performed RNA immunoprecipitation (RIP) assays to detect Ess2-interacting snRNAs. We found that Ess2 associated with U6 snRNA as well as U1 and U4 snRNAs. Experiments using Ess2 deletion mutants showed that a C-terminus deletion mutant of Ess2 (1-399 a. a.) lost its ability to associate with snRNAs, whereas the N-terminus domain of Ess2 (1-200 a. a.) associated with Rorγ/γt, but not with snRNAs. Interestingly, experiments using anti-ROR common antibody showed that Rors also associated with U4 and U6 snRNAs. Ess2 knockdown in a T cell hybridoma (68-41 cells) abrogated the interaction between spliceosomes and Rors. An Ess2-dependent association was also found between an lncRNA (Rmrp) and Rors. We thus propose that Ess2 associates with both transcriptional factors and spliceosomal complexes and modulates splicing reactions coupled with transcription factors.


Subject(s)
Nuclear Proteins/metabolism , Nuclear Receptor Subfamily 1, Group F, Member 3/metabolism , RNA, Small Nuclear/metabolism , Spliceosomes/metabolism , Animals , Cell Line , Gene Knockdown Techniques , HEK293 Cells , Humans , Mice , Mutation , Nuclear Proteins/analysis , Nuclear Proteins/genetics , Nuclear Receptor Subfamily 1, Group F, Member 3/genetics , Protein Interaction Domains and Motifs , RNA Splicing , RNA, Small Nuclear/genetics , Spliceosomes/genetics , Transcriptional Activation
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