title: Identification of a novel lipin homologue from the parasitic protozoan Trypanosoma brucei.
abstract: BACKGROUND
  Arginine methylation is a post-translational modification that expands the functional diversity of proteins. Kinetoplastid parasites contain a relatively large group of protein arginine methyltransferases (PRMTs) compared to other single celled eukaryotes. Several T. brucei proteins have been shown to serve as TbPRMT substrates in vitro, and a great number of proteins likely to undergo methylation are predicted by the T. brucei genome. This indicates that a large number of proteins whose functions are modulated by arginine methylation await discovery in trypanosomes. Here, we employed a yeast two-hybrid screen using as bait the major T. brucei type I PRMT, TbPRMT1, to identify potential substrates of this enzyme.
  
  
  RESULTS
  We identified a protein containing N-LIP and C-LIP domains that we term TbLpn. These domains are usually present in a family of proteins known as lipins, and involved in phospholipid biosynthesis and gene regulation. Far western and co-immunoprecipitation assays confirmed the TbPRMT1-TbLpn interaction. We also demonstrated that TbLpn is localized mainly to the cytosol, and is methylated in vivo. In addition, we showed that, similar to mammalian and yeast proteins with N-LIP and C-LIP domains, recombinant TbLpn exhibits phosphatidic acid phosphatase activity, and that two conserved aspartic acid residues present in the C-LIP domain are critical for its enzymatic activity.
  
  
  CONCLUSIONS
  This study reports the characterization of a novel trypanosome protein and provides insight into its enzymatic activity and function in phospholipid biosynthesis. It also indicates that TbLpn functions may be modulated by arginine methylation.
authors: Department of Biology, The College at Brockport, State University of New York, Brockport, NY, USA. mpelleti@brockport.edu
authors: Michel
authors: M
authors: Pelletier
authors: Alyssa S
authors: AS
authors: Frainier
authors: Dominic N
authors: DN
authors: Munini
authors: Jenna M
authors: JM
authors: Wiemer
authors: Amber R
authors: AR
authors: Karpie
authors: Jeff J
authors: JJ
authors: Sattora
mesh_terms: D026481:Blotting, Far-Western; D006801:Humans; D047468:Immunoprecipitation; D010711:Phosphatidate Phosphatase / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism*; D015800:Protozoan Proteins / Q000302:isolation & purification / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000201:enzymology*; D020798:Two-Hybrid System Techniques
chemical_list: D015800:Protozoan Proteins; C512613:PRMT1 protein, Trypanosoma brucei; D011484:Protein-Arginine N-Methyltransferases; D010711:Phosphatidate Phosphatase
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