DeAngelo Joseph D, Maron Maxim I, Roth Jacob S, Silverstein Aliza M, Gupta Varun, Stransky Stephanie, Basken Joel, Azofeifa Joey, Sidoli Simone, Gamble Matthew J, Shechter David
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
Contributed equally.
bioRxiv. 2024 Aug 12:2024.08.09.607355. doi: 10.1101/2024.08.09.607355.
Protein Arginine Methyltransferase 5 (PRMT5) regulates RNA splicing and transcription by symmetric dimethylation of arginine residues (Rme2s/SDMA) in many RNA binding proteins. However, the mechanism by which PRMT5 couples splicing to transcriptional output is unknown. Here, we demonstrate that a major function of PRMT5 activity is to promote chromatin escape of a novel, large class of mRNAs that we term Genomically Retained Incompletely Processed Polyadenylated Transcripts (GRIPPs). Using nascent and total transcriptomics, spike-in controlled fractionated cell transcriptomics, and total and fractionated cell proteomics, we show that PRMT5 inhibition and knockdown of the PRMT5 SNRP (Sm protein) adapter protein pICln (CLNS1A) -but not type I PRMT inhibition-leads to gross detention of mRNA, SNRPB, and SNRPD3 proteins on chromatin. Compared to most transcripts, these chromatin-trapped polyadenylated RNA transcripts have more introns, are spliced slower, and are enriched in detained introns. Using a combination of PRMT5 inhibition and inducible isogenic wildtype and arginine-mutant SNRPB, we show that arginine methylation of these snRNPs is critical for mediating their homeostatic chromatin and RNA interactions. Overall, we conclude that a major role for PRMT5 is in controlling transcript processing and splicing completion to promote chromatin escape and subsequent nuclear export.
蛋白质精氨酸甲基转移酶5(PRMT5)通过对许多RNA结合蛋白中的精氨酸残基进行对称二甲基化(Rme2s/SDMA)来调节RNA剪接和转录。然而,PRMT5将剪接与转录输出偶联的机制尚不清楚。在这里,我们证明PRMT5活性的一个主要功能是促进一类新的、大量的mRNA从染色质上逃逸,我们将这类mRNA称为基因组保留的未完全加工的多聚腺苷酸化转录本(GRIPPs)。使用新生转录组学和总转录组学、掺入对照的分级细胞转录组学以及总细胞和分级细胞蛋白质组学,我们表明PRMT5抑制以及PRMT5的小核核糖核蛋白(Sm蛋白)衔接蛋白pICln(CLNS1A)的敲低——而不是I型PRMT抑制——导致mRNA、小核核糖核蛋白B(SNRPB)和小核核糖核蛋白D3(SNRPD3)蛋白在染色质上的大量滞留。与大多数转录本相比,这些被染色质捕获的多聚腺苷酸化RNA转录本有更多的内含子,剪接速度更慢,并且在滞留的内含子中富集。使用PRMT5抑制与诱导型同基因野生型和精氨酸突变型SNRPB相结合的方法,我们表明这些小核核糖核蛋白的精氨酸甲基化对于介导它们的稳态染色质和RNA相互作用至关重要。总体而言,我们得出结论,PRMT5的一个主要作用是控制转录本加工和剪接完成,以促进染色质逃逸和随后的核输出。