Here we provide evidence that EBNA2 is methylated in vivo and that methylation of EBNA2 is a prerequisite for binding to SMN. We present SMN as a novel binding partner of EBNA2 by showing that EBNA2 colocalizes with SMN in nuclear gems and that both proteins can be coimmunoprecipitated from cellular extract. Furthermore, in vitro methylation of either wild-type EBNA2 or a glutathione S-transferase-EBNA2 fusion protein encompassing the arginine-glycine (RG) repeat element is necessary for in vitro binding to the Tudor domain of SMN. The recently shown functional cooperation of SMN and EBNA2 in transcriptional activation and the previous observation of a severely reduced transformation potential yet strongly enhanced transcriptional activity of an EBNA2 mutant lacking the RG repeat indicate that binding of SMN to EBNA2 is a critical step in B-cell transformation by Epstein-Barr virus.The Epstein-Barr virus (EBV) causes infectious mononucleosis and is associated with a variety of human tumors (reviewed in reference 25). The virus transforms B cells into continuously growing lymphoblastoid cell lines by expressing 12 genes (reviewed in reference 1). The nuclear antigen EBNA2 is the first viral gene expressed after infection and is essential for EBVmediated transformation through the activation of viral and cellular genes like the viral EBNAs and the latent membrane proteins LMP1, LMP2A, and LMP2B as well as the cellular CD21, CD23, c-fgr, and c-myc (reviewed in reference 13) and AML-2 genes (26). EBNA2 does not bind directly but is tethered to promoters by interacting with cellular transcription factors like RBPJ, Sp-1/Spi-B, hnRNP-D/AUF1, or ATF/ CRE. Domains of EBNA2 critical for transformation of B cells and for activation of gene expression had been identified previously through mutational analysis (6,28,29). EBNA2 binds to RBPJ with its conserved WWP 325 (18) motif, the deletion of which results in a severe reduction of activation of the LMP1 promoter and a complete loss of transforming capacity (6). An adjacent arginine-glycine (RG) repeat element between amino acids (aa) 337 and 354 of EBNA2 was shown elsewhere to be critical but not essential for B-cell transformation in vitro, but the deletion of aa 337 to 354 increased the potential of EBNA2 to activate the LMP1 promoter (29). While most interacting partners bind to the C terminus of EBNA2, we had recently demonstrated that EBNA2 interacts through its N-terminal aa 121 to 216 with a putative helicase/ATPase termed DP103 (12), which in turn binds to SMN, the product of the spinal muscular atrophy gene (SMN) (4,5,30). The survival of motor neurons (SMN) gene is lost or mutated in spinal muscular atrophy (SMA) (17). SMN is found in different macromolecular complexes, and one of these has recently been shown to facilitate the assembly of spliceosomal U snRNPs by mediating the attachment of the Sm proteins onto snRNAs U1, U2, U4, and U5 (7,20,22). The function of the SMN complex involves direct binding of Sm proteins to SMN, an interaction that is strongly enha...

Epstein-Barr virus nuclear antigen 2 (EBNA2) is essential for viral transformation of B cells and transactivates cellular and viral target genes by binding RBPJ tethered to cognate promoter elements. EBNA2interacts with the DEAD-box protein DP103 (DDX20/Gemin3), which in turn is complexed to the survival motor neuron (SMN) protein. SMN is implicated in RNA processing, but a role in transcriptional regulation has also been suggested. Here, we show that DP103 and SMN are complexed in B cells and that SMN coactivates the viral LMP promoter in the presence of EBNA2 in reporter gene assays and in vivo. Subcellular localization studies revealed that nuclear gems and/or coiled bodies containing DP103 and SMN are targeted by EBNA2. Protein-protein interaction experiments demonstrated that DP103 binds to SMN exon 6 and that both EBNA2 and SMN interact with the C terminus of DP103. Furthermore, a DP103 binding-deficient SMN mutant was released from nuclear gems and/or coiled bodies and further enhanced coactivation. In addition, impaired transactivation of a DP103 binding-deficient EBNA2 mutant was rescued by overexpression of SMN. Testing different promoter constructs in luciferase assays showed that RBPJ is required but not sufficient for coactivation by EBNA2 and SMN. Overall, our data suggest that EBNA2 might target spliceosomal complexes by binding to DP103, thereby releasing SMN which subsequently exerts a coactivational function within the RNA-polymerase II transcription complex on the LMP1 promoter.The Epstein-Barr virus (EBV) causes infectious mononucleosis and is linked to the genesis of several human lymphoproliferative diseases (for a review, see reference 33). The EBVencoded nuclear antigen 2 (EBNA2) is a viral transactivator essential for EBV-induced transformation of resting human B lymphocytes, by promoting the expression of the transforming latent membrane proteins LMP1 and 2, the nuclear EBV Cp promoter-driven EBNA proteins, and the cellular genes CD23 and c-fgr (for review, see reference 15). EBNA2 does not bind directly to DNA but exerts its function by interacting with the cellular proteins RBPJ (CBF1) and, on the more complex LMP1 promoter, also Spi1 (PU.1), tethered to cognate response elements (12,17,20,45,46). Transcriptional activation is induced by binding of the C-terminal acidic domain (5) to components of the basal RNA polymerase II transcription machinery, such as RPA70, TAF40, TFIIB, and TFIIH (38, 39), and recruitment of the coactivators p300, CBP, and PCAF histone deacetylase (14, 41). In addition, by attracting the hSWI/SNF complex (42, 43) and targeting histone H1 (9, 34), EBNA2 likely promotes relief of nucleosome-mediated gene repression.We have recently shown that EBNA2 binds to DP103, a novel member of the DEAD-box family of putative RNA helicases (10). DP103 is a ubiquitously expressed 103-kDa phosphoprotein with an RNA-dependent ATPase activity; its other functions, in particular with regard to its interaction with EBNA2, remained unknown. While the work presented here was in prog...

Insulin-resistant skeletal muscle of ZDF rats is characterised by a specific gene expression profile with increased levels of Scd1. An insulin-resistant phenotype similar to that obtained by treatment with palmitate and high glucose can be induced in vitro by overexpression of SCD1 in muscle cells. This supports the hypothesis that elevated SCD1 expression is a possible cause of insulin resistance and type 2 diabetes.

No abstract

PIONEER (Prostate Cancer DIagnOsis and TreatmeNt Enhancement through the power of big data in EuRope) is a European network of excellence for big data in prostate cancer, consisting of 32 private and public stakeholders from 9 countries across Europe. Launched by the Innovative Medicines Initiative 2 and part of the Big Data for Better Outcomes Programme (BD4BO), the overarching goal of PIONEER is to provide high-quality evidence on prostate cancer management by unlocking the potential of big data. The project has identified critical evidence gaps in prostate cancer care, via a detailed prioritisation exercise including all key stakeholders. By standardising and integrating existing high quality and multidisciplinary data sources from prostate cancer patients across different stages of the disease, rich big data will be assembled into a single innovative data platform for research. Based on a unique set of methodologies, PIONEER aims at advancing the field of prostate cancer care with particular focus on improving prostate cancer-related outcomes, health system efficiency by streamlining patient management, and the quality of health and social care delivered to all prostate cancer patients and their families. The literature suggests there is underuse of effective treatments and overuse of ineffective treatment. For example, androgen deprivation therapy is sometimes overused in situations where it is not recommended. It is therefore crucial to identify the best treatment option for the individual patient.