Type I and II PRMTs inversely regulate post-transcriptional intron detention through Sm and CHTOP methylation

eLife

Maxim I Maron¹  Simone Sidoli¹  Jacob S Roth¹  David Shechter¹  Charles C Query²  Varun Gupta²  Alyssa D Casill³  Matthew J Gamble⁴ 

[1] Department of Biochemistry, Albert Einstein College of Medicine, Bronx, United States;Department of Cell Biology, Albert Einstein College of Medicine, Bronx, United States;Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, United States;Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, United States;Department of Cell Biology, Albert Einstein College of Medicine, Bronx, United States;

关键词: PRMT5;    PRMT1;    retained detained introns;    RNA processing;    snRNP;    A549;    Detained introns;    DI;    Mass spectrometry;    PTMs;    Post-translational modifications;    Transcription;    SDMA;    MMA;    ADMA;    Rme2s;    Rme2a;    Rme1;    PRMTs;    PRMT1;    PRMT5;    CARM1;    PRMT4;    Protein Arginine Methyltransferases;    Splicing;    Post-transcriptional processing;   

DOI  :  10.7554/eLife.72867

来源: eLife Sciences Publications, Ltd

【 摘 要 】

Protein arginine methyltransferases (PRMTs) are required for the regulation of RNA processing factors. Type I PRMT enzymes catalyze mono- and asymmetric dimethylation; Type II enzymes catalyze mono- and symmetric dimethylation. To understand the specific mechanisms of PRMT activity in splicing regulation, we inhibited Type I and II PRMTs and probed their transcriptomic consequences. Using the newly developed Splicing Kinetics and Transcript Elongation Rates by Sequencing (SKaTER-seq) method, analysis of co-transcriptional splicing demonstrated that PRMT inhibition resulted in altered splicing rates. Surprisingly, co-transcriptional splicing kinetics did not correlate with final changes in splicing of polyadenylated RNA. This was particularly true for retained introns (RI). By using actinomycin D to inhibit ongoing transcription, we determined that PRMTs post-transcriptionally regulate RI. Subsequent proteomic analysis of both PRMT-inhibited chromatin and chromatin-associated polyadenylated RNA identified altered binding of many proteins, including the Type I substrate, CHTOP, and the Type II substrate, SmB. Targeted mutagenesis of all methylarginine sites in SmD3, SmB, and SmD1 recapitulated splicing changes seen with Type II PRMT inhibition, without disrupting snRNP assembly. Similarly, mutagenesis of all methylarginine sites in CHTOP recapitulated the splicing changes seen with Type I PRMT inhibition. Examination of subcellular fractions further revealed that RI were enriched in the nucleoplasm and chromatin. Taken together, these data demonstrate that, through Sm and CHTOP arginine methylation, PRMTs regulate the post-transcriptional processing of nuclear, detained introns.

【 授权许可】

CC BY