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

Mol Biochem Parasitol. 2001 Nov;118(1):49-59. doi: 10.1016/s0166-6851(01)00367-x.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Arginine / metabolism*
  • Mass Spectrometry
  • Methylation
  • Mitochondria / metabolism*
  • Protein Processing, Post-Translational*
  • Protein-Arginine N-Methyltransferases / metabolism
  • Protozoan Proteins*
  • RNA Editing
  • RNA, Guide / genetics
  • RNA, Guide / metabolism*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Trypanosoma brucei brucei / genetics
  • Trypanosoma brucei brucei / growth & development
  • Trypanosoma brucei brucei / metabolism*

Substances

  • Protozoan Proteins
  • RBP16 protein, Trypanosoma
  • RNA, Guide
  • RNA-Binding Proteins
  • Arginine
  • Protein-Arginine N-Methyltransferases