Functional cooperation of Epstein-Barr virus nuclear antigen 2 and the survival motor neuron protein in transactivation of the viral LMP1 promoter.
Epstein-Barr virus nuclear antigen 2 (EBNA2) is essential for viral transformation of B cells and transactivates cellular and viral target genes by binding RBPJkappa tethered to cognate promoter elements. EBNA2 interacts 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 RBPJkappa 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.
Voss
M D
MD
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universitätskliniken, 66421 Homburg/Saar, Germany.
Hille
A
A
Barth
S
S
Spurk
A
A
Hennrich
F
F
Holzer
D
D
Mueller-Lantzsch
N
N
Kremmer
E
E
Grässer
F A
FA
D001483:Base Sequence; D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein; D017931:DNA Primers; D019309:Epstein-Barr Virus Nuclear Antigens / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D009419:Nerve Tissue Proteins / Q000502:physiology*; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins; D015533:Transcriptional Activation / Q000502:physiology*; D014763:Viral Matrix Proteins / Q000235:genetics*; D014764:Viral Proteins
D017362:Cyclic AMP Response Element-Binding Protein; D017931:DNA Primers; C402641:EBNA-2 protein, Human herpesvirus 4; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins