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The RS/RGG Domain of the SR‐like Protein Npl3 is Required for Efficient RNA Splicing in Yeast
Author(s) -
Lin Mengjia M,
Gala Kinisha,
Lobby Jenna,
Moazamian Aron,
Kress Tracy L
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.459.7
Subject(s) - rna splicing , sr protein , splicing factor , exonic splicing enhancer , alternative splicing , biology , rna , microbiology and biotechnology , rna binding protein , gene , genetics , protein domain , exon , computational biology
Npl3 is an SR‐like protein found in Saccharomyces cerevisiae that functions in co‐transcriptional pre‐mRNA splicing by promoting the recruitment of splicing factors to the pre‐mRNA during transcription1. It contains two RNA recognition motifs (RRMs), an RS/RGG domain rich in arginine and serine repeats, and an uncharacterized N‐terminus. While Npl3 binds RNA2–4 via the RRM domains5, the role of each of the functional domains in splicing is currently unknown. Here we have generated a panel of flag‐tagged Npl3 proteins lacking one or more of the protein domains. We identified genetic interactions between Npl3 mutations lacking the RS/RGG domain and mutations in genes that encode RNA splicing factors, suggesting that the RS/RGG domain is important for RNA splicing. Indeed, we show that deletion of the RS/RGG domain of Npl3 causes a significant decrease in RNA splicing efficiency of Npl3‐dependent pre‐mRNAs. In addition, deletion of the RS/RGG domain abolishes the ability of Npl3 to interact with splicing factors, which strongly suggest that the RS/RGG domain is required to recruit the splicing machinery during transcription. Interestingly, we found that phosphorylation of the RS/RGG domain is not important for RNA splicing. Replacing the Npl3 RS/RGG domain with the RS/RGG domain from the Candida albicans Npl3 ortholog6 restores growth, interaction with splicing proteins, and RNA splicing. Together this work demonstrates that the RS/RGG domain is required for the activity of Npl3 in RNA splicing. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .