Arginine methylation of a mitochondrial guide RNA binding protein from Trypanosoma brucei

Michel Pelletier, Ye Xu, Xu Wang, Sotir Zahariev, Sandor Pongor, John M. Aletta, Laurie K. Read

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

RBP16 is a mitochondrial Y-box protein from the parasitic protozoan Trypanosoma brucei that binds guide RNAs and ribosomal RNAs. It is comprised of an N-terminal cold-shock domain and a C-terminal domain rich in glycine and arginine residues, resembling the RGG RNA-binding motif. Arginine residues found within RGG domains are frequently asymmetrically dimethylated by a class of enzymes termed protein arginine methyltransferases (PRMTs). As Arg-93 of RBP16 exists in the context of a preferred sequence for asymmetric arginine dimethylation (G/FGGRGGG/F), we investigated whether modified arginines are present in native RBP16 by MALDI-TOF and post-source decay analyses. These analyses confirmed that Arg-93 is dimethylated. In addition, Arg-78 exists as an unmodified or as a monomethylated derivative, and Arg-85 is present in forms corresponding to the unmodified, di-, and tri-methylated state. While Arg-93 is apparently constitutively dimethylated, the methylation of Arg-78 and Arg-85 is mutually exclusive. Furthermore, whole cell extracts from procyclic form T. brucei are able to methylate bacterially expressed RBP16 (rRBP16), as well as endogenous proteins, in the presence of S-adenosyl-L-[methyl-3H]methionine. While assays of mitochondrial extracts suggest a small amount of PRMT may also be present in this subcellular compartment, the majority of trypanosome PRMT activity is extramitochondrial. We show that rRBP16 is methylated in trypanosome extracts through the action of a type I methyltransferase as well as serving as a substrate for heterologous mammalian type I PRMTs. In addition, we demonstrate the presence of type II PRMT activity in trypanosome cell extracts. These results suggest that protein arginine methylation is a common posttranslational modification in trypanosomes, and that it may regulate the function of RBP16.

Original languageEnglish (US)
Pages (from-to)49-59
Number of pages11
JournalMolecular and Biochemical Parasitology
Volume118
Issue number1
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Guide RNA
Protein-Arginine N-Methyltransferases
Trypanosoma brucei brucei
RNA-Binding Proteins
Trypanosomiasis
Methylation
Arginine
Cell Extracts
Protozoan Proteins
Ribosomal RNA
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Methyltransferases
Post Translational Protein Processing
Glycine
Shock
Proteins
mitochondrial RNA
Enzymes

Keywords

  • Arginine methylation
  • Guide RNA
  • RNA binding protein
  • RNA editing
  • Trypanosoma

ASJC Scopus subject areas

  • Molecular Biology
  • Parasitology

Cite this

Arginine methylation of a mitochondrial guide RNA binding protein from Trypanosoma brucei. / Pelletier, Michel; Xu, Ye; Wang, Xu; Zahariev, Sotir; Pongor, Sandor; Aletta, John M.; Read, Laurie K.

In: Molecular and Biochemical Parasitology, Vol. 118, No. 1, 2001, p. 49-59.

Research output: Contribution to journalArticle

Pelletier, Michel ; Xu, Ye ; Wang, Xu ; Zahariev, Sotir ; Pongor, Sandor ; Aletta, John M. ; Read, Laurie K. / Arginine methylation of a mitochondrial guide RNA binding protein from Trypanosoma brucei. In: Molecular and Biochemical Parasitology. 2001 ; Vol. 118, No. 1. pp. 49-59.
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T1 - Arginine methylation of a mitochondrial guide RNA binding protein from Trypanosoma brucei

AU - Pelletier, Michel

AU - Xu, Ye

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AU - Zahariev, Sotir

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AU - Aletta, John M.

AU - Read, Laurie K.

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AB - RBP16 is a mitochondrial Y-box protein from the parasitic protozoan Trypanosoma brucei that binds guide RNAs and ribosomal RNAs. It is comprised of an N-terminal cold-shock domain and a C-terminal domain rich in glycine and arginine residues, resembling the RGG RNA-binding motif. Arginine residues found within RGG domains are frequently asymmetrically dimethylated by a class of enzymes termed protein arginine methyltransferases (PRMTs). As Arg-93 of RBP16 exists in the context of a preferred sequence for asymmetric arginine dimethylation (G/FGGRGGG/F), we investigated whether modified arginines are present in native RBP16 by MALDI-TOF and post-source decay analyses. These analyses confirmed that Arg-93 is dimethylated. In addition, Arg-78 exists as an unmodified or as a monomethylated derivative, and Arg-85 is present in forms corresponding to the unmodified, di-, and tri-methylated state. While Arg-93 is apparently constitutively dimethylated, the methylation of Arg-78 and Arg-85 is mutually exclusive. Furthermore, whole cell extracts from procyclic form T. brucei are able to methylate bacterially expressed RBP16 (rRBP16), as well as endogenous proteins, in the presence of S-adenosyl-L-[methyl-3H]methionine. While assays of mitochondrial extracts suggest a small amount of PRMT may also be present in this subcellular compartment, the majority of trypanosome PRMT activity is extramitochondrial. We show that rRBP16 is methylated in trypanosome extracts through the action of a type I methyltransferase as well as serving as a substrate for heterologous mammalian type I PRMTs. In addition, we demonstrate the presence of type II PRMT activity in trypanosome cell extracts. These results suggest that protein arginine methylation is a common posttranslational modification in trypanosomes, and that it may regulate the function of RBP16.

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