Asymmetric Arginine dimethylation of Epstein–Barr virus nuclear antigen 2 promotes DNA targeting

• Published in: Virology (New York, N.Y.) Vol. 397; no. 2; pp. 299 - 310
• Main Authors: Gross, Henrik, Barth, Stephanie, Palermo, Richard D, Mamiani, Alfredo, Hennard, Christine, Zimber-Strobl, Ursula, West, Michelle J, Kremmer, Elisabeth, Grässer, Friedrich A
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.02.2010
• Subjects: 60 APPLIED LIFE SCIENCES; aDMA; AMINO ACIDS; ANIMAL CELLS; Animals; ANTIGENS; ARGININE; Arginine - metabolism; ASYMMETRY; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CARBOXYLIC ACIDS; Cell Line; CHEMICAL REACTIONS; CONNECTIVE TISSUE CELLS; DNA; DNA - metabolism; DNA-binding; EBNA2; EBV; Epstein-Barr virus; Epstein-Barr Virus Nuclear Antigens - metabolism; Gene Expression; GENES; Herpesvirus 4, Human - physiology; Humans; Immunoglobulin J Recombination Signal Sequence-Binding Protein - metabolism; Infectious Disease; INHIBITION; LEUKOCYTES; LYMPHOCYTES; MATERIALS; METHYLATION; Mice; Mice, Inbred BALB C; MICROORGANISMS; Nuclear antigen 2; NUCLEIC ACIDS; ONCOGENIC VIRUSES; ORGANIC ACIDS; ORGANIC COMPOUNDS; PARASITES; Promoter Regions, Genetic; PROMOTERS; Protein Binding; PROTEINS; Rats; RBPJκ; RESIDUES; sDMA; SOMATIC CELLS; TRANSCRIPTION FACTORS; Viral Matrix Proteins - biosynthesis; Viral Proteins - metabolism; VIRUSES; sDMA; RBPJκ; Arginine; EBV; EBNA2; aDMA; Methylation; Epstein–Barr virus; Nuclear antigen 2; DNA-binding
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2009.11.023

Abstract The Epstein–Barr virus (EBV) growth-transforms B-lymphocytes. The virus-encoded nuclear antigen 2 (EBNA2) is essential for transformation and activates gene expression by association with DNA-bound transcription factors such as RBPJκ (CSL/CBF1). We have previously shown that EBNA2 contains symmetrically dimethylated Arginine (sDMA) residues. Deletion of the RG-repeat results in a reduced ability of the virus to immortalise B-cells. We now show that the RG repeat also contains asymmetrically dimethylated Arginines (aDMA) but neither non-methylated (NMA) Arginines nor citrulline residues. We demonstrate that only aDMA-containing EBNA2 is found in a complex with DNA-bound RBPJκ in vitro and preferentially associates with the EBNA2-responsive EBV C, LMP1 and LMP2A promoters in vivo . Inhibition of methylation in EBV-infected cells results in reduced expression of the EBNA2-regulated viral gene LMP1, providing additional evidence that methylation is a prerequisite for DNA-binding by EBNA2 via association with the transcription factor RBPJκ.