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Protein Arginine Methyltransferases: Promising Targets for Cancer Therapy Jee Won Hwang1,Yenacho1,Gyu-Unbae1,Su-Namkim2 and Yong Kee Kim 1

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Protein Arginine Methyltransferases: Promising Targets for Cancer Therapy Jee Won Hwang1,Yenacho1,Gyu-Unbae1,Su-Namkim2 and Yong Kee Kim 1 Hwang et al. Experimental & Molecular Medicine (2021) 53:788–808 https://doi.org/10.1038/s12276-021-00613-y Experimental & Molecular Medicine REVIEW ARTICLE Open Access Protein arginine methyltransferases: promising targets for cancer therapy Jee Won Hwang1,YenaCho1,Gyu-UnBae1,Su-NamKim2 and Yong Kee Kim 1 Abstract Protein methylation, a post-translational modification (PTM), is observed in a wide variety of cell types from prokaryotes to eukaryotes. With recent and rapid advancements in epigenetic research, the importance of protein methylation has been highlighted. The methylation of histone proteins that contributes to the epigenetic histone code is not only dynamic but is also finely controlled by histone methyltransferases and demethylases, which are essential for the transcriptional regulation of genes. In addition, many nonhistone proteins are methylated, and these modifications govern a variety of cellular functions, including RNA processing, translation, signal transduction, DNA damage response, and the cell cycle. Recently, the importance of protein arginine methylation, especially in cell cycle regulation and DNA repair processes, has been noted. Since the dysregulation of protein arginine methylation is closely associated with cancer development, protein arginine methyltransferases (PRMTs) have garnered significant interest as novel targets for anticancer drug development. Indeed, several PRMT inhibitors are in phase 1/2 clinical trials. In this review, we discuss the biological functions of PRMTs in cancer and the current development status of PRMT inhibitors in cancer therapy. 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; G G Introduction ω-N ,N -asymmetric dimethyl arginine (ADMA), and G G Since the discovery of arginine residue methylation on ω-N ,N’ -symmetric dimethyl arginine (SDMA)8. PRMTs histone proteins1, protein arginine methylation has been are classified into three subgroups based on the type of emphasized as an indispensable post-translational mod- methyl arginine they produce: Type I PRMTs (PRMT1, 2, ification (PTM) and an epigenetic regulation mechan- 3, 4, 6, and 8) generate MMA and ADMA, Type II PRMTs ism2,3. Arginine methylation is catalyzed by a family of (PRMT5 and 9) produce MMA and SDMA, and Type III enzymes called protein arginine methyltransferases PRMT (PRMT7) produces only MMA7,9. (PRMTs), and nine PRMTs have been identified in The arginine residue consists of a guanidino group on mammals to date (Fig. 1a)2,4,5. All PRMTs share four its side chain, which is protonated and positively charged conserved sequence motifs (I, post-I, II, and III) and one at physiological pH3,5. The guanidino group forms mul- THW loop, which compose the S-adenosyl-L-methionine tiple hydrogen bonds that bind with other interacting (AdoMet) binding pocket in the tertiary structure6,7. proteins or cofactors2,5. Although the methylated arginine PRMTs transfer a methyl group from the AdoMet residue retains its positive charge, the ability to form molecule to the guanidino group of the arginine residue in hydrogen bonds is reduced, probably affecting the substrate proteins8. There are three types of methyl protein-protein interaction. In addition, arginine methy- G arginine (Fig. 1b): ω-N -monomethyl arginine (MMA), lation is very stable compared to that of other PTMs, and hence, its kinetics are less dynamic2,5. PRMTs are asso- ciated with many essential cellular processes, including Correspondence: Yong Kee Kim ([email protected]) transcription, splicing, translation, signal transduction, 1Research Institute of Pharmaceutical Sciences, College of Pharmacy, ’ DNA damage and repair, and cell cycle regulation Sookmyung Women s University, Seoul 04310, Republic of Korea 2–4 2Natural Product Research Institute, Korea Institute of Science and Technology, (Fig. 2) , and the knockout phenotypes of some PRMTs Gangneung 25451, Republic of Korea © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a linktotheCreativeCommons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Official journal of the Korean Society for Biochemistry and Molecular Biology Hwang et al. Experimental & Molecular Medicine (2021) 53:788–808 789 (a) (b) Name Domain structureClassificaon Cellular localizaon arginine 1ab c d e 371 NH + NH PRMT1 type I cytoplasm/nucleus 2 2 50 361 C 1ab c d e 433 PRMT2 SH3 type I cytoplasm/nucleus NH 30 89 99 432 (CH2)3 1ab c d e 531 PRMT3 Zn-F type I cytoplasm Protein 48 71 271 531 1 608 ab c d e type I, II, and III CARM1 TAD type I cytoplasm/nucleus 146 453 499 608 1ab c d e 637 asymmetric di-methyl mono-methyl symmetric di-methyl PRMT5 TIM-barrel type II cytoplasm/nucleus arginine (ADMA) arginine (MMA) arginine (SDMA) 13 292 308 615 1ab c d e 375 PRMT6 type I nucleus CH CH CH 3 3 3 CH3 44 374 + + + NH2 N NH2 NH NH NH 1 692 ab c d e ab c d CH PRMT7 type III cytoplasm/nucleus C 3 C C 14 345 358 684 NH type I NH type II NH SH3 binding mof 1 394 ab c d e (CH ) (CH ) (CH ) PRMT8 myr type I plasma membrane 2 3 2 3 2 3 73 394 TPR Protein Protein Protein 1ab c d e ab c d e 845 PRMT9 type II cytoplasm 137 466 530 845 Fig. 1 Protein arginine methylation and responsible enzymes. a The mammalian PRMT family. Nine PRMTs were identified, and these have unique signatures (dark blue lines) with high sequence similarity (a, Motif I: VLD/EVGXGXG; b, Post-I: V/IXG/AXD/E; c, Motif II: F/I/VDI/L/K; d, Motif III: LR/KXXG; e, THW loop). Their enzymatic types and cellular localization are shown. b Types of arginine methylation. The arginine residue has two equivalent nitrogen atoms in its guanidino group. Types I, II, and III PRMTs generate monomethyl arginine (MMA) marks. The subsequent generation of asymmetric dimethyl arginine (ADMA) is catalyzed by type I enzymes (PRMT1, PRMT2, PRMT3, CARM1, PRMT6, and PRMT8), and symmetric dimethyl arginine (SDMA) is produced by type II enzymes (PRMT5 and PRMT9). PRMT7, a type III enzyme, generates only MMA. – show embryonic or perinatal lethality2,10 12, indicating the arginine residues not only serve as key epigenetic marks significance of PRMTs in maintaining functional home- but also engage in crosstalk with other epigenetic ostasis in biological systems. Tissue-specific deletion marks21,22. These orchestrated epigenetic modifications studies of PRMTs strongly support the supposition that are recognized by epigenetic reader proteins, leading to they are involved in cancer and metabolic, immune, the recruitment of activating/repressing transcriptional neurodegenerative, and muscular disorders4,13,14. Since machinery. The histone modifications generated by the dysregulation of PRMTs has been closely associated PRMTs and their roles are summarized in Table 1. The with cancer development2,15,16, the use of PRMTs as novel methylation status of an arginine residue in histones can targets for anticancer drug development is rapidly determine whether the transcription process is activated increasing. Recent studies have revealed considerable or suppressed. For example, H4R3me2a, a modification advances in the identification of clinically relevant PRMT generated by PRMT1/PRMT3, acts as a mark of activated inhibitors17,18. Here, we focus on the biological functions transcription, whereas H4R3me2s, generated by PRMT5, of PRMTs in cancer and the therapeutic potential of functions as a repression mark, implying that there is a PRMT inhibitors. sophisticated and competitive mechanism between PRMTs for regulating the transcription process. In addi- Biological functions of protein arginine tion to histone proteins, various proteins involved in methylation transcription, such as transcription factors, coactivators, As histone proteins tightly regulate gene transcription and corepressors, are also methylated by PRMTs (Table 1 through various PTMs, including acetylation, lysine and Fig. 2)23. Hence, PRMTs also contribute to the precise methylation, phosphorylation, ubiquitination, and regulation of the transcription process. A number of SUMOylation19,20, early studies of PRMTs have also RNA-binding proteins (RBPs) have RG/RGG-rich motifs focused on their epigenetic functions. PRMTs synthesize that have been established as representative consensus methyl arginine on nucleosomes after being recruited into sequences of PRMTs24,25. Indeed, theoretical insights and chromatin remodeling complexes, and these methylated proteomic analysis revealed that several RBPs are Official journal of the Korean Society for Biochemistry and Molecular Biology Hwang et al. Experimental & Molecular Medicine (2021) 53:788–808 790 Genec Mutaons Diet Aging Hormones PRMTs Environments MMA, ADMA, SDMA Histones Non-histones H3: R2me2a(s), R8me2a(s), R17me2a, R26me2a, C/EBPα, FOXO1, GLI1/2, Sm proteins, hnRNPs, R42me2a EGFR, CRAF, p65/RelA, Smad6/7, INCENP, CDK4, H4: R3me2a(s) H2A: R29me2a p53, RUVBL1, 53BP1, BRCA1,
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