In vitro and in vivo analysis of the major type I protein arginine methyltransferase from Trypanosoma brucei

Pelletier, M; Pasternack, Deborah A.; Read, Laurie K.

doi:10.1016/j.molbiopara.2005.08.015

Cited by 31 publications

(83 citation statements)

References 57 publications

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“…We previously reported that TbPRMT1 was unable to methylate RBP16 in vitro (Pelletier et al 2005), while another mitochondrial RGGcontaining protein, TBRGG1, was robustly methylated. We postulated that a cofactor such as RNA or the RBP16-associated protein, p22 (Hayman et al 2001), might be required for RBP16 methylation.…”

Section: Rbp16 Is a Substrate For Tbprmt1 In Vitromentioning

confidence: 98%

“…We previously described the creation and initial characterization of a clonal procyclic form TbPRMT1 RNAi cell line (Pelletier et al 2005). In the previous studies, we demonstrated that although growth was unaffected, total cellular asymmetric dimethylarginine was dramatically reduced in these cells on day 4 following tet induction of RNAi.…”

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 98%

“…TbPRMT1 is the trypanosome homolog of the mammalian PRMT1 and yeast HMT1 enzymes, and constitutes the major type I PRMT in T. brucei (Pelletier et al 2005). We previously described the creation and initial characterization of a clonal procyclic form TbPRMT1 RNAi cell line (Pelletier et al 2005).…”

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 99%

“…In the previous studies, we demonstrated that although growth was unaffected, total cellular asymmetric dimethylarginine was dramatically reduced in these cells on day 4 following tet induction of RNAi. To determine whether protein arginine methylation has an impact on mitochondrial gene expression in trypanosomes, we first examined mitochondrial RNA metabolism following tet-induced depletion of TbPRMT1 (Pelletier et al 2005). Total RNA was isolated from TbPRMT1 RNAi cells either induced for 4 d in the presence of 2.5 mg/mL tet, or left uninduced prior to RNA isolation.…”

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 99%

“…We postulated that a cofactor such as RNA or the RBP16-associated protein, p22 (Hayman et al 2001), might be required for RBP16 methylation. In an attempt to identify conditions under which RBP16 is a substrate for TbPRMT1, we performed in vitro methylation assays as previously described (Pelletier et al 2005) with slight modifications to the enzyme preparation (see Materials and Methods). His-RBP16 (Miller and Read 2003) was incubated with MBP-TbPRMT1 (Pelletier et al 2005) in the presence of the methyl donor [ 3 H]AdoMet for 18 h at 27 o C. As shown in Figure 2, there are two bands present in the MBP-PRMT1/His-RBP16 (lane 2), a faint band corresponding to full-length His-RBP16 (arrow) and a more prominent band that presumably corresponds to a FIGURE 1.…”

Section: Rbp16 Is a Substrate For Tbprmt1 In Vitromentioning

confidence: 99%

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Arginine methylation regulates mitochondrial gene expression in Trypanosoma brucei through multiple effector proteins

Goulah¹,

Pelletier²,

Read³

2006

RNA

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Arginine methylation is a post-translational modification that impacts gene expression in both the cytoplasm and nucleus. Here, we demonstrate that arginine methylation also affects mitochondrial gene expression in the protozoan parasite, Trypanosoma brucei. Down-regulation of the major trypanosome type I protein arginine methyltransferase, TbPRMT1, leads to destabilization of specific mitochondrial mRNAs. We provide evidence that some of these effects are mediated by the mitochondrial RNAbinding protein, RBP16, which we previously demonstrated affects both RNA editing and stability. TbPRMT1 catalyzes methylation of RBP16 in vitro. Further, MALDI-TOF-MS analysis of RBP16 isolated from TbPRMT1-depleted cells indicates that, in vivo, TbPRMT1 modifies two of the three known methylated arginine residues in RBP16. Expression of mutated, nonmethylatable RBP16 in T. brucei has a dominant negative effect, leading to destabilization of a subset of those mRNAs affected by TbPRMT1 depletion. Our results suggest that the specificity and multifunctional nature of RBP16 are due, at least in part, to the presence of differentially methylated forms of the protein. However, some effects of TbPRMT1 depletion on mitochondrial gene expression cannot be accounted for by RBP16 action. Thus, these data implicate additional, unknown methylproteins in mitochondrial gene regulation.

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Section: Rbp16 Is a Substrate For Tbprmt1 In Vitromentioning

confidence: 98%

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 98%

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 99%

Section: Tbprmt1 Depletion Reveals a Role For Arginine Methylation Inmentioning

confidence: 99%

Section: Rbp16 Is a Substrate For Tbprmt1 In Vitromentioning

confidence: 99%

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Arginine methylation regulates mitochondrial gene expression in Trypanosoma brucei through multiple effector proteins

Goulah¹,

Pelletier²,

Read³

2006

RNA

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Functional interplay between protein arginine methyltransferases in Trypanosoma brucei

Lott

Fisk

et al. 2014

MicrobiologyOpen

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Arginine methylation is a common posttranslational modification that has far-reaching cellular effects. Trypanosoma brucei is an early-branching eukaryote with four characterized protein arginine methyltransferases (PRMTs), one additional putative PRMT, and over 800 arginine methylated proteins, suggesting that arginine methylation has widespread impacts in this organism. While much is known about the activities of individual T. brucei PRMTs (TbPRMTs), little is known regarding how TbPRMTs function together in vivo. In this study, we analyzed single and selected double TbPRMT knockdowns for the impact on expression of TbPRMTs and global methylation status. Repression of TbPRMT1 caused a decrease in asymmetric dimethylarginine and a marked increase in monomethylarginine that was catalyzed by TbPRMT7, suggesting that TbPRMT1 and TbPRMT7 can compete for the same substrate. We also observed an unexpected and strong interdependence between TbPRMT1 and TbPRMT3 protein levels. This finding, together with the observation of similar methyl landscape profiles in TbPRMT1 and TbPRMT3 repressed cells, strongly suggests that these two enzymes form a functional complex. We show that corepression of TbPRMT6/7 synergistically impacts growth of procyclic-form T. brucei. Our findings also implicate the actions of noncanonical, and as yet unidentified, PRMTs in T. brucei. Together, our studies indicate that TbPRMTs display a functional interplay at multiple levels.

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Type I and II PRMTs regulate catabolic as well as detoxifying processes in Aspergillus nidulans

Bauer

Lechner

Pidroni

et al. 2019

Fungal Genetics and Biology

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In filamentous fungi, arginine methylation has been implicated in morphogenesis, mycotoxin biosynthesis, pathogenicity, and stress response although the exact role of this posttranslational modification in these processes remains obscure. Here, we present the first genome-wide transcriptome analysis in filamentous fungi that compared expression levels of genes regulated by type I and type II protein arginine methyltransferases (PRMTs). In Aspergillus nidulans, three conserved type I and II PRMTs are present that catalyze asymmetric or symmetric dimethylation of arginines. We generated a double type I mutant (ΔrmtA/rmtB) and a combined type I and type II mutant (ΔrmtB/rmtC) to perform genome-wide comparison of their effects on gene expression, but also to monitor putative overlapping activities and reciprocal regulations of type I and type II PRMTs in Aspergillus. Our study demonstrates, that rmtA and rmtC as type I and type II representatives act together as repressors of proteins that are secreted into the extracellular region as the majority of up-regulated genes are mainly involved in catabolic pathways that constitute the secretome of Aspergillus. In addition to a strong up-regulation of secretory genes we found a significant enrichment of down-regulated genes involved in processes related to oxidation-reduction, transmembrane transport and secondary metabolite biosynthesis. Strikingly, nearly 50% of down-regulated genes in both double mutants correspond to redox reaction/oxidoreductase processes, a remarkable finding in light of our recently observed oxidative stress phenotypes of ΔrmtA and ΔrmtC. Finally, analysis of nuclear and cytoplasmic extracts for mono-methylated proteins revealed the presence of both, common and specific substrates of RmtA and RmtC. Thus, our data indicate that type I and II PRMTs in Aspergillus seem to co-regulate the same biological processes but also specifically affect other pathways in a non-redundant fashion.

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In vitro and in vivo analysis of the major type I protein arginine methyltransferase from Trypanosoma brucei

Cited by 31 publications

References 57 publications

Arginine methylation regulates mitochondrial gene expression in Trypanosoma brucei through multiple effector proteins

Arginine methylation regulates mitochondrial gene expression in Trypanosoma brucei through multiple effector proteins

Functional interplay between protein arginine methyltransferases in Trypanosoma brucei

Type I and II PRMTs regulate catabolic as well as detoxifying processes in Aspergillus nidulans

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