Protein Arginine Methylation in Parasitic Protozoa

ABSTRACT Protozoa constitute the earliest branch of the eukaryotic lineage, and several groups of protozoans are serious parasites of humans and other animals. Better understanding of biochemical pathways that are either in common with or divergent from those of higher eukaryotes is integral in the defense against these parasites. In yeast and humans, the posttranslational methylation of arginine residues in proteins affects myriad cellular processes, including transcription, RNA processing, DNA replication and repair, and signal transduction. The protein arginine methyltransferases (PRMTs) that catalyze these reactions, which are unique to the eukaryotic kingdom of organisms, first become evident in protozoa. In this review, we focus on the current understanding of arginine methylation in multiple species of parasitic protozoa, including Trichomonas , Entamoeba , Toxoplasma , Plasmodium , and Trypanosoma spp., and discuss how arginine methylation may play important and unique roles in each type of parasite. We mine available genomic and transcriptomic data to inventory the families of PRMTs in different parasites and the changes in their abundance during the life cycle. We further review the limited functional studies on the roles of arginine methylation in parasites, including epigenetic regulation in Apicomplexa and RNA processing in trypanosomes. Interestingly, each of the parasites considered herein has significantly differing sets of PRMTs, and we speculate on the importance of this diversity in aspects of parasite biology, such as differentiation and antigenic variation.

Download Full-text

Characterization of the Drosophila protein arginine methyltransferases DART1 and DART4

Biochemical Journal ◽

10.1042/bj20031176 ◽

2004 ◽

Vol 379 (2) ◽

pp. 283-289 ◽

Cited By ~ 47

Author(s):

Marie-Chloé BOULANGER ◽

Tina Branscombe MIRANDA ◽

Steven CLARKE ◽

Marco di FRUSCIO ◽

Beat SUTER ◽

...

Keyword(s):

Rna Binding ◽

Developmental Stages ◽

Rna Binding Proteins ◽

Genetic System ◽

Arginine Methylation ◽

Type I ◽

Protein Arginine Methyltransferases ◽

Arginine Residues ◽

Drosophila Protein

The role of arginine methylation in Drosophila melanogaster is unknown. We identified a family of nine PRMTs (protein arginine methyltransferases) by sequence homology with mammalian arginine methyltransferases, which we have named DART1 to DART9 (Drosophilaarginine methyltransferases 1–9). In keeping with the mammalian PRMT nomenclature, DART1, DART4, DART5 and DART7 are the putative homologues of PRMT1, PRMT4, PRMT5 and PRMT7. Other DART family members have a closer resemblance to PRMT1, but do not have identifiable homologues. All nine genes are expressed in Drosophila at various developmental stages. DART1 and DART4 have arginine methyltransferase activity towards substrates, including histones and RNA-binding proteins. Amino acid analysis of the methylated arginine residues confirmed that both DART1 and DART4 catalyse the formation of asymmetrical dimethylated arginine residues and they are type I arginine methyltransferases. The presence of PRMTs in D. melanogaster suggest that flies are a suitable genetic system to study arginine methylation.

Download Full-text

Methylation of the nuclear poly(A)-binding protein by type I protein arginine methyltransferases – how and why

Biological Chemistry ◽

10.1515/hsz-2013-0121 ◽

2013 ◽

Vol 394 (8) ◽

pp. 1029-1043 ◽

Cited By ~ 3

Author(s):

Elmar Wahle ◽

Bodo Moritz

Keyword(s):

Active Sites ◽

Binding Protein ◽

Arginine Methylation ◽

Fine Tuning ◽

General Mechanism ◽

Type I ◽

Protein Arginine Methyltransferases ◽

Low Protein ◽

Backbone Conformation ◽

Arginine Residues

Abstract Asymmetric dimethylation of arginine side chains in proteins is a frequent posttranslational modification, catalyzed by type I protein arginine methyltransferases (PRMTs). This article summarizes what is known about this modification in the nuclear poly(A)-binding protein (PABPN1). PABPN1 contains 13 dimethylated arginine residues in its C-terminal domain. Three enzymes, PRMT1, 3, and 6, can methylate PABPN1. Although 26 methyl groups are transferred to one PABPN1 molecule, the PRMTs do so in a distributive reaction, i.e., only a single methyl group is transferred per binding event. As PRMTs form dimers, with the active sites accessible from a small central cavity, backbone conformation around the methyl-accepting arginine is an important determinant of substrate specificity. Neither the association of PABPN1 with poly(A) nor its role in poly(A) tail synthesis is affected by arginine methylation. At least at low protein concentration, methylation does not affect the protein’s tendency to oligomerize. The dimethylarginine residues of PABPN1 are located in the binding site for its nuclear import receptor, transportin. Arginine methylation weakens this interaction about 10-fold. Very recent evidence suggests that arginine methylation as a way of fine-tuning the interactions between transportin and its cargo may be a general mechanism.

Download Full-text

Recruitment of the NineTeen Complex to the activated spliceosome requires AtPRMT5

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1522458113 ◽

2016 ◽

Vol 113 (19) ◽

pp. 5447-5452 ◽

Cited By ~ 14

Author(s):

Xian Deng ◽

Tiancong Lu ◽

Lulu Wang ◽

Lianfeng Gu ◽

Jing Sun ◽

...

Keyword(s):

Molecular Mechanism ◽

Normal Growth ◽

Arginine Methylation ◽

Mrna Splicing ◽

Sm Proteins ◽

Protein Arginine Methyltransferases ◽

Small Nuclear Ribonucleoprotein ◽

Spliceosome Activation ◽

Multiple Species ◽

Nuclear Ribonucleoprotein

Protein arginine methylation, catalyzed by protein arginine methyltransferases (PRMTs), is involved in a multitude of biological processes in eukaryotes. Symmetric arginine dimethylation mediated by PRMT5 modulates constitutive and alternative pre-mRNA splicing of diverse genes to regulate normal growth and development in multiple species; however, the underlying molecular mechanism remains largely unknown. A genetic screen for suppressors of an Arabidopsis symmetric arginine dimethyltransferase mutant, atprmt5, identified two gain-of-function alleles of pre-mRNA processing factor 8 gene (prp8-8 and prp8-9), the highly conserved core component of the U5 small nuclear ribonucleoprotein (snRNP) and the spliceosome. These two atprmt5 prp8 double mutants showed suppression of the developmental and splicing alterations of atprmt5 mutants. In atprmt5 mutants, the NineTeen complex failed to be assembled into the U5 snRNP to form an activated spliceosome; this phenotype was restored in the atprmt5 prp8-8 double mutants. We also found that loss of symmetric arginine dimethylation of Sm proteins prevents recruitment of the NineTeen complex and initiation of spliceosome activation. Together, our findings demonstrate that symmetric arginine dimethylation has important functions in spliceosome assembly and activation, and uncover a key molecular mechanism for arginine methylation in pre-mRNA splicing that impacts diverse developmental processes.

Download Full-text

Prmt7 is dispensable in tissue culture models for adipogenic differentiation

F1000Research ◽

10.12688/f1000research.2-279.v1 ◽

2013 ◽

Vol 2 ◽

pp. 279 ◽

Cited By ~ 4

Author(s):

Yu-Jie Hu ◽

Saïd Sif ◽

Anthony N. Imbalzano

Keyword(s):

Gene Expression ◽

Tissue Culture ◽

Adipogenic Differentiation ◽

Arginine Methylation ◽

Type Ii ◽

Protein Arginine Methyltransferases ◽

Over Expression ◽

Core Histones ◽

Culture Models ◽

Arginine Residues

Protein arginine methylation is a common posttranslational modification that has been implicated in numerous biological processes including gene expression. The mammalian genome encodes nine protein arginine methyltransferases (Prmts) that catalyze monomethylation, asymmetric dimethylation, and symmetric dimethylation on arginine residues. Protein arginine methyltransferase 7 (Prmt7) is categorized as a type II and type III enzyme that produces symmetric dimethylated arginine and monomethylated arginine, respectively. However, the biological role of Prmt7 is not well characterized. We previously showed that Prmt5, a type II Prmt that associates with Brg1-based SWI/SNF chromatin remodeling complex, is required for adipocyte differentiation. Since Prmt7 also associates with Brg1-based SWI/SNF complex and modifies core histones, we hypothesized that Prmt7 might play a role in transcriptional regulation of adipogenesis. In the present study, we determined that the expression of Prmt7 did not change throughout adipogenic differentiation of C3H10T1/2 mesenchymal cells. Knockdown or over-expression of Prmt7 had no effect on lipid accumulation or adipogenic gene expression in differentiating C3H10T1/2 cells or in C/EBPα-reprogrammed NIH3T3 fibroblasts. Based on these results, we conclude that Prmt7, unlike Prmt5, is dispensable for adipogenic differentiation in tissue culture models.

Download Full-text

Functional study of Leishmania braziliensis protein arginine methyltransferases (PRMTs) reveals that PRMT1 and PRMT5 are required for macrophage infection

10.1101/2021.09.22.461376 ◽

2021 ◽

Author(s):

Lucas Lorenzon ◽

Jose Carlos Quilles ◽

Gustavo Daniel Campagnaro ◽

Leticia Almeida ◽

Flavio Protasio Veras ◽

...

Keyword(s):

Gene Expression ◽

Life Cycle ◽

Rna Binding ◽

Arginine Methylation ◽

Functional Study ◽

Leishmania Braziliensis ◽

Macrophage Infection ◽

Protein Arginine Methyltransferases ◽

Life Cycle Stages ◽

Arginine Residues

In trypanosomatids, regulation of gene expression occurs mainly at the posttranscriptional level, and RNA-binding proteins (RBPs) are key players in determining the fates of transcripts. RBPs are major targets of protein arginine methyltransferases (PRMTs), which posttranslationally regulate the RNA-binding capacity and other macromolecular interactions of RBPs by transferring methyl groups to protein arginine residues. Herein, we present the results of a study that functionally characterized the five predicted PRMTs in Leishmania braziliensis by gene knockout and endogenous protein HA tagging using CRISPR/Cas9 gene editing. We report that arginine methylation profiles vary among Leishmania species and that target protein methylation changes across different L. braziliensis life cycle stages, with higher PRMT expression in the promastigote stages than in the axenic amastigote stage. Knockout of some of the L. braziliensis PRMTs led to significant changes in global arginine methylation patterns without affecting promastigote axenic growth. Deletion of either PRMT1 or PRMT3 disrupted most type I PRMT activity, resulting in a global increase in monomethyl arginine (MMA) levels, which is mainly catalyzed by PRMT7. Putative targets and/or PRMT-interacting proteins were identified by coimmunoprecipitation using HA-tagged PRMTs, revealing a network of target RBPs and suggesting functional interactions between them and a relevant participation in epigenetic control of gene expression. Finally, we demonstrate that L. braziliensis PRMT1 and PRMT5 are required for efficient macrophage infection in vitro, and that in the absence of PRMT1 and PRMT5, axenic amastigote proliferation is impaired. The results indicate that arginine methylation is modulated across life cycle stages in L. braziliensis and show possible functional overlap and cooperation among the different PRMTs in targeting proteins. Overall, our data suggest important regulatory roles of these proteins throughout the L. braziliensis life cycle, showing that arginine methylation is important for parasite-host cell interactions.

Download Full-text

Scintillation Proximity Assay of Arginine Methylation

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057111414903 ◽

2011 ◽

Vol 17 (2) ◽

pp. 237-244 ◽

Cited By ~ 25

Author(s):

Jiang Wu ◽

Nan Xie ◽

You Feng ◽

Y. George Zheng

Keyword(s):

High Throughput Screening ◽

Scintillation Counter ◽

Regulation Of Gene Expression ◽

Arginine Methylation ◽

Activity Measurement ◽

Histone H4 ◽

Protein Arginine Methyltransferases ◽

Scintillation Proximity Assay ◽

Assay Procedure ◽

Arginine Residues

Methylation of arginine residues, catalyzed by protein arginine methyltransferases (PRMTs), is one important protein posttranslational modification involved in epigenetic regulation of gene expression. A fast and effective assay for PRMT can provide valuable information for dissecting the biological functions of PRMTs, as well as for screening small-molecule inhibitors of arginine methylation. Currently, among the methods used for PRMT activity measurement, many contain laborious separation procedures, which restrict the applications of these assays for high-throughput screening (HTS) in drug discovery. The authors report here a mix-and-measure method to measure PRMT activity based on the principle of scintillation proximity assay (SPA). In this assay, 3H-AdoMet was used as methyl donor, and biotin-modified histone H4 peptide served as a methylation substrate. Following the methylation reaction catalyzed by PRMTs, streptavidin-coated SPA beads were added to the reaction solution, and SPA signals were detected by a MicroBeta scintillation counter. No separation step is needed, which simplifies the assay procedure and greatly enhances the assay speed. Particularly, the miniaturization and robustness suggest that this method is suited for HTS of PRMT inhibitors.

Download Full-text

PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of Neisseria gonorrhoeae

mSphere ◽

10.1128/msphere.00562-19 ◽

2019 ◽

Vol 4 (5) ◽

Cited By ~ 1

Author(s):

Egon A. Ozer ◽

Lauren L. Prister ◽

Shaohui Yin ◽

Billy H. Ward ◽

Stanimir Ivanov ◽

...

Keyword(s):

Neisseria Gonorrhoeae ◽

Antigenic Variation ◽

Amplicon Sequencing ◽

Common Mechanism ◽

Macrophage Infection ◽

Gene Encoding ◽

Transmitted Infection ◽

Variable Regions ◽

Content Type ◽

Strain Fa1090

ABSTRACT Gene diversification is a common mechanism pathogens use to alter surface structures to aid in immune avoidance. Neisseria gonorrhoeae uses a gene conversion-based diversification system to alter the primary sequence of the gene encoding the major subunit of the pilus, pilE. Antigenic variation occurs when one of the nonexpressed 19 silent copies donates part of its DNA sequence to pilE. We have developed a method using Pacific Biosciences (PacBio) amplicon sequencing and custom software to determine pilin antigenic variation frequencies. The program analyzes 37 variable regions across the strain FA1090 1-81-S2 pilE gene and can be modified to determine sequence variation from other starting pilE sequences or other diversity generation systems. Using this method, we measured pilin antigenic variation frequencies for various derivatives of strain FA1090 and showed we can also analyze pilin antigenic variation frequencies during macrophage infection. IMPORTANCE Diversity generation systems are used by many unicellular organism to provide subpopulations of cell with different properties that are available when needed. We have developed a method using the PacBio DNA sequencing technology and a custom computer program to analyze the pilin antigenic variation system of the organism that is the sole cause of the sexually transmitted infection, gonorrhea.

Download Full-text

Are STATS Arginine-methylated?

Journal of Biological Chemistry ◽

10.1074/jbc.c400606200 ◽

2005 ◽

Vol 280 (23) ◽

pp. 21700-21705 ◽

Cited By ~ 52

Author(s):

Waraporn Komyod ◽

Uta-Maria Bauer ◽

Peter C. Heinrich ◽

Serge Haan ◽

Iris Behrmann

Keyword(s):

Arginine Methylation ◽

Signaling Cascades ◽

Post Translational Modification ◽

Protein Arginine Methyltransferases ◽

Stat Proteins ◽

Fibrosarcoma Cells ◽

Catalytically Active ◽

Interferon Resistance ◽

Stat Pathway

Transcription factors of the STAT (signal transducer and activator of transcription) family are important in signal transduction of cytokines. They are subject to post-translational modification by phosphorylation on tyrosine and serine residues. Recent evidence suggested that STATs are methylated on a conserved arginine residue within the N-terminal region. STAT arginine methylation has been described to be important for STAT function and loss of arginine methylation was discussed to be involved in interferon resistance of cancer cells. Here we provide several independent lines of evidence indicating that the issue of arginine methylation of STATs has to be reassessed. First, we show that treatment of melanoma and fibrosarcoma cells with inhibitors used to suppress methylation (N-methyl-2-deoxyadenosine, adenosine, dl-homocysteine) had profound and rapid effects on phosphorylation of STAT1 and STAT3 but also on p38 and Erk signaling cascades which are known to cross-talk with the Jak/STAT pathway. Second, we show that anti-methylarginine antibodies did not precipitate specifically STAT1 or STAT3. Third, we show that mutation of Arg31 to Lys led to destabilization of STAT1 and STAT3, implicating an important structural role of Arg31. Finally, purified catalytically active protein arginine methyltransferases (PRMT1, -2, -3, -4, and -6) did not methylate STAT proteins, and cotransfection with PRMT1 did not affect STAT1-controlled reporter gene activity. Taken together, our data suggest the absence of arginine methylation of STAT1 and STAT3.

Download Full-text

Favourite puzzles

English Text Construction ◽

10.1075/etc.10.2.01van ◽

2017 ◽

Vol 10 (2) ◽

pp. 187-198 ◽

Cited By ~ 1

Author(s):

Lieven Vandelanotte

Keyword(s):

Cognitive Linguistics ◽

Literary Discourse ◽

Special Issue ◽

Content Type ◽

Doctoral Research ◽

Functional Studies

This introduction to the special issue “Grammar, usage and discourse: Functional studies offered to Kristin Davidse” first briefly reviews Kristin Davidse’s rich and varied trajectory in functional and cognitive linguistics, highlighting in particular the links between the domains represented by the contributions to the issue and the doctoral research she has supervised over the years. The central questions surrounding grammar (especially interpersonal grammar), usage and discourse (including literary discourse) which inform the different contributions are subsequently discussed, and a concluding section offers a number of celebratory and grateful salutes.

Download Full-text

Epigenetic control via allosteric regulation of mammalian protein arginine methyltransferases

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1706978114 ◽

2017 ◽

Vol 114 (38) ◽

pp. 10101-10106 ◽

Cited By ~ 21

Author(s):

Kanishk Jain ◽

Cyrus Y. Jin ◽

Steven G. Clarke

Keyword(s):

Cancer Metastasis ◽

Gene Activation ◽

Kinetic Studies ◽

Arginine Methylation ◽

Histone H4 ◽

Protein Arginine Methyltransferases ◽

Wide Range ◽

Substrate Concentrations

Arginine methylation on histones is a central player in epigenetics and in gene activation and repression. Protein arginine methyltransferase (PRMT) activity has been implicated in stem cell pluripotency, cancer metastasis, and tumorigenesis. The expression of one of the nine mammalian PRMTs, PRMT5, affects the levels of symmetric dimethylarginine (SDMA) at Arg-3 on histone H4, leading to the repression of genes which are related to disease progression in lymphoma and leukemia. Another PRMT, PRMT7, also affects SDMA levels at the same site despite its unique monomethylating activity and the lack of any evidence for PRMT7-catalyzed histone H4 Arg-3 methylation. We present evidence that PRMT7-mediated monomethylation of histone H4 Arg-17 regulates PRMT5 activity at Arg-3 in the same protein. We analyzed the kinetics of PRMT5 over a wide range of substrate concentrations. Significantly, we discovered that PRMT5 displays positive cooperativity in vitro, suggesting that this enzyme may be allosterically regulated in vivo as well. Most interestingly, monomethylation at Arg-17 in histone H4 not only raised the general activity of PRMT5 with this substrate, but also ameliorated the low activity of PRMT5 at low substrate concentrations. These kinetic studies suggest a biochemical explanation for the interplay between PRMT5- and PRMT7-mediated methylation of the same substrate at different residues and also suggest a general model for regulation of PRMTs. Elucidating the exact relationship between these two enzymes when they methylate two distinct sites of the same substrate may aid in developing therapeutics aimed at reducing PRMT5/7 activity in cancer and other diseases.

Download Full-text