Arginine methylation of RNA‐binding proteins regulates cell function and differentiation

Blackwell, Ernest; Ceman, Stephanie

doi:10.1002/mrd.22024

Cited by 79 publications

(85 citation statements)

References 155 publications

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“…7D, lanes 8 to 11 and 13). The latter observation confirms our previous finding that the combinatorial change of arginines 22,23,25, and 26 to alanine changes pUL69 into a UAP56-binding-deficient mutant (6,8,9). Interestingly, however, the arginines at positions 28 and 29 were not required for UAP56 interaction, since a mutant harboring the respective substitutions was coprecipitated by Myc-UAP56 (Fig.…”

Section: Resultssupporting

confidence: 80%

“…As suggested by the varying amino acid sequences and different subcellular localizations, PRMTs have distinct substrate specificities (22). In contrast to PMKTs, which mainly modify histones, PRMTs predominantly target nonhistone substrates, for instance, transcription factors, various RNAprocessing factors like hnRNPs, spliceosomal Sm proteins, the DEAD box helicase VASA, and RNA-binding proteins involved in neuronal development, such as FMRP, HuR, and HuD (23). Importantly, arginine methylation was demonstrated to modulate the nucleocytoplasmic transport and the activity of various RNAprocessing factors (24)(25)(26)(27)(28).…”

Section: Importancementioning

confidence: 99%

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pUL69 of Human Cytomegalovirus Recruits the Cellular Protein Arginine Methyltransferase 6 via a Domain That Is Crucial for mRNA Export and Efficient Viral Replication

Thomas

Sonntag

Müller

et al. 2015

J Virol

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The regulatory protein pUL69 of human cytomegalovirus acts as a viral mRNA export factor, facilitating the cytoplasmic accumulation of unspliced RNA via interaction with the cellular mRNA export factor UAP56. Here we provide evidence for a posttranslational modification of pUL69 via arginine methylation within the functionally important N terminus. First, we demonstrated a specific immunoprecipitation of full-length pUL69 as well as pUL69aa1-146 by a mono/dimethylarginine-specific antibody. Second, we observed a specific electrophoretic mobility shift upon overexpression of the catalytically active protein arginine methyltransferase 6 (PRMT6). Third, a direct interaction of pUL69 and PRMT6 was confirmed by yeast two-hybrid and coimmunoprecipitation analyses. We mapped the PRMT6 interaction motif to the pUL69 N terminus and identified critical amino acids within the arginine-rich R1 box of pUL69 that were crucial for PRMT6 and/or UAP56 recruitment. In order to test the impact of putative methylation substrates on the functions of pUL69, we constructed various pUL69 derivatives harboring arginine-to-alanine substitutions and tested them for RNA export activity. Thus, we were able to discriminate between arginines within the R1 box of pUL69 that were crucial for UAP56/PRMT6-interaction and/or mRNA export activity. Remarkably, nuclear magnetic resonance (NMR) analyses revealed the same ␣-helical structures for pUL69 sequences encoding either the wild type R1/R2 boxes or a UAP56/PRMT6 binding-deficient derivative, thereby excluding the possibility that R/A amino acid substitutions within R1 affected the secondary structure of pUL69. We therefore conclude that the pUL69 N terminus is methylated by PRMT6 and that this critically affects the functions of pUL69 for efficient mRNA export and replication of human cytomegalovirus. IMPORTANCEThe UL69 protein of human cytomegalovirus is a multifunctional regulatory protein that acts as a viral RNA export factor with a critical role for efficient replication. Here, we demonstrate that pUL69 is posttranslationally modified via arginine methylation and that the protein methyltransferase PRMT6 mediates this modification. Furthermore, arginine residues with a crucial function for RNA export and for binding of the cellular RNA export factor UAP56 as well as PRMT6 were mapped within the arginine-rich R1 motif of pUL69. Importantly, we demonstrated that mutation of those arginines did not alter the secondary structure of R1, suggesting that they may serve as critical methylation substrates. In summary, our study reveals a novel posttranslational modification of pUL69 which has a significant impact on the function of this important viral regulatory protein. Since PRMTs appear to be amenable to selective inhibition by small molecules, this may constitute a novel target for antiviral therapy. E very mammalian or avian herpesvirus encodes a member of the ICP27 protein family which functions as a posttranscriptional activator by facilitating the nuclear export of intronless mRNAs ...

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Section: Resultssupporting

confidence: 80%

Section: Importancementioning

confidence: 99%

pUL69 of Human Cytomegalovirus Recruits the Cellular Protein Arginine Methyltransferase 6 via a Domain That Is Crucial for mRNA Export and Efficient Viral Replication

Thomas

Sonntag

Müller

et al. 2015

J Virol

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“…In addition, FMRP has been shown to undergo two post-translational modifications: phosphorylation and arginine methylation , Stetler et al 2006. It has been suggested that phosphorylation might modulate FMRP properties such as association with actively translating polyribosomes and with Dicer (Ceman et al 2003, Cheever & Ceman 2009) while methylation affects both its protein-protein and protein-RNA interactions (Denman 2002, Dolzhanskaya et al 2006, Stetler et al 2006, Blackwell et al 2010, Blackwell & Ceman 2012. The presence of a nuclear localization signal and a nuclear export signal suggests shuttling between the nucleus and the cytoplasm (Eberhart et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Expression of fragile X mental retardation protein and Fmr1 mRNA during folliculogenesis in the rat

et al. 2013

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Fragile X mental retardation protein (FMRP) belongs to a small family of RNA-binding proteins. Its absence or inactivity is responsible for fragile X syndrome, the most common cause of inherited mental retardation. Despite its ubiquitous expression, FMRP function and expression remain almost understudied in non-neuronal tissues, though previous studies on germline development during oogenesis may suggest a special function of this protein also in ovarian tissue. In addition, the well-documented association of FMR1 premutation state with fragile X-related premature ovarian insufficiency adds interest to the role of FMRP in ovarian physiology. The aim of the present work was to investigate the expression of Fmr1 mRNA and its protein, FMRP, at different stages of rat follicular development. By immunohistochemical studies we demonstrated FMRP expression in granulosa, theca and germ cells in all stages of follicular development. In addition, changes in Fmr1 expression, both at the protein and mRNA levels, were observed. FMRP levels increased upon follicular development while preantral and early antral follicles presented similar levels of Fmr1 transcripts with decreased expression in preovulatory follicles. These observations suggest that Fmr1 expression in the ovary is regulated at different and perhaps independent levels. In addition, our results show expression of at least four different isoforms of FMRP during all stages of follicular growth with expression patterns that differ from those observed in brain and testis. Our study shows a regulated expression of Fmr1, both at mRNA and protein levels, during rat follicular development.

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“…Arginine methylation is an important post-translational modification that may occur as symmetrical dimethylarginines (sDMAs), asymmetrical dimethylarginines (aDMAs), or monomethylarginines (Krause et al 2007;Bedford and Clarke 2009;Blackwell and Ceman 2012). sDMAs occur in "sDMA motifs" composed of multiple arginine-glycine (RG) and arginine-alanine (RA) repeats in various proteins, and modulate diverse cellular processes.…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial protein BmPAPI modulates the length of mature piRNAs

Honda

Kirino

Maragkakis

et al. 2013

RNA

140

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PIWI proteins and their associated PIWI-interacting RNAs (piRNAs) protect genome integrity by silencing transposons in animal germlines. The molecular mechanisms and components responsible for piRNA biogenesis remain elusive. PIWI proteins contain conserved symmetrical dimethylarginines (sDMAs) that are specifically targeted by TUDOR domain-containing proteins. Here we report that the sDMAs of PIWI proteins play crucial roles in PIWI localization and piRNA biogenesis in Bombyx mori-derived BmN4 cells, which harbor fully functional piRNA biogenesis machinery. Moreover, RNAi screenings for Bombyx genes encoding TUDOR domain-containing proteins identified BmPAPI, a Bombyx homolog of Drosophila PAPI, as a factor modulating the length of mature piRNAs. BmPAPI specifically recognized sDMAs and interacted with PIWI proteins at the surface of the mitochondrial outer membrane. BmPAPI depletion resulted in 3 ′ -terminal extensions of mature piRNAs without affecting the piRNA quantity. These results reveal the BmPAPI-involved piRNA precursor processing mechanism on mitochondrial outer membrane scaffolds.

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Arginine methylation of RNA‐binding proteins regulates cell function and differentiation

Cited by 79 publications

References 155 publications

pUL69 of Human Cytomegalovirus Recruits the Cellular Protein Arginine Methyltransferase 6 via a Domain That Is Crucial for mRNA Export and Efficient Viral Replication

pUL69 of Human Cytomegalovirus Recruits the Cellular Protein Arginine Methyltransferase 6 via a Domain That Is Crucial for mRNA Export and Efficient Viral Replication

Expression of fragile X mental retardation protein and Fmr1 mRNA during folliculogenesis in the rat

Mitochondrial protein BmPAPI modulates the length of mature piRNAs

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