Asymmetric Arginine dimethylation of Epstein-Barr virus nuclear antigen 2 promotes DNA targeting.
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 RBPJkappa (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 RBPJkappa 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 RBPJkappa.
Gross
Henrik
H
Institut für Virologie, Haus 47, Universitätsklinikum, 66421 Homburg/Saar, Germany.
Barth
Stephanie
S
Palermo
Richard D
RD
Mamiani
Alfredo
A
Hennard
Christine
C
Zimber-Strobl
Ursula
U
West
Michelle J
MJ
Kremmer
Elisabeth
E
Grässer
Friedrich A
FA
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism*; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein / Q000378:metabolism*; D008745:Methylation; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D014763:Viral Matrix Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000378:metabolism*
C402641:EBNA-2 protein, Human herpesvirus 4; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; C497570:RBPJ protein, human; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D004247:DNA; D001120:Arginine