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

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 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.

EBV nuclear antigen EBNALP dismisses transcription repressors NCoR and RBPJ from enhancers and EBNA2 increases NCoR-deficient RBPJ DNA binding

“…Alternatively, EBNA2 amino acids 292-310, which are important for RBPJ and SKIP interaction, and EBNA2 amino acids 318-322, which interact directly with the RBPJ β trefoil domain (BTD) (21,22,26,49) may together exclude NCoR from RBPJ-EBNA2 complexes or inhibit NCoR antibody immunoprecipitation. Recently, arginine dimethylated EBNA2 has been reported to interact more strongly with DNA, perhaps selectively through NCoR-deficient RBPJ (50). Putative RBPJ inhibition of NCoR immunoprecipitation is unlikely, given the otherwise positive correlation of NCoR and RBP ChIP data.…”

The NP9 protein encoded by the human endogenous retrovirus HERV‐K(HML‐2) negatively regulates gene activation of the Epstein‐Barr virus nuclear antigen 2 (EBNA2)

“…34 The antibodies 6F12 and 13B10 which recognise the symmetrically and asymmetrically dimethylated Arginine-Glycine repeat of EBNA2, respectively, and the GST-specific monoclonal antibody 6G9 have been described recently. 9 Precipitated EBNA2 and GST-NP9 were detected using R3 and 6G9, respectively.…”

“…Electroporation of Raji cells with pSG5-NP9 was carried out as described earlier 9 . The expression of NP9 was enhanced by treatment of the cells with TPA for 18 hr.…”

“…8 We have recently shown that the RG-repeat, in addition to the symmetrically dimethylated Arginines (SDMA) also contains asymmetrically dimethylated Arginine residues (ADMA), which are preferentially found in DNA-bound EBNA2. 9 The human endogenous retroviruses (HERV) encompass $8% of the human genome and represent mostly defective proviruses. 10 Some members of the HERV-K family contain complete open reading frames for viral gag, pol and env genes.…”

Symmetric Dimethylarginine as Predictor of Graft Loss and All-Cause Mortality in Renal Transplant Recipients.

“…A high proportion of cancers in RTR are lymphoproliferative and related to infections with Epstein-Barr virus, human herpesvirus 8, human papilloma virus or the hepatitis B and C viruses (41). Symmetric dimethylarginine residuals are shown to be important constituents of the Epstein-Barr virusYencoded nuclear antigen 2 which is responsible for growth transformations in B lymphocytes (42,43), but whether virus-related malignancy is associated with increased plasma levels of SDMA is not known. Further studies are needed to elaborate on this as well as to investigate whether associations between SDMA and specific non-CV causes of death can be found in other populations.…”