Dissecting the contribution of EBNA3C domains important for EBV-induced B-cell growth and proliferation

Oncotarget
Hem Chandra JhaErle S Robertson

Abstract

Epstein-Barr virus (EBV) is an oncogenic gammaherpes virus which is linked to pathogenesis of several human lymphatic malignancies. The EBV essential latent antigen EBNA3C is critical for efficient conversion of primary human B-lymphocytes to lymphoblastic cell lines and for continued LCL growth. EBNA3C, an EBV latent antigen with oncogenic potential can bind and regulate the functions of a wide range of cellular transcription factors. In our current reverse genetics study, we deleted the full length EBNA3C, and independently the RBP-Jκ and Nm23-H1 binding sites within EBNA3C using BACmid recombinant engineering methodology. Our experiments demonstrated that deletion of the EBV EBNA3C open reading frame (ORF) and more specifically the residues 621-675 which binds Nm23H1 and SUMO-1 showed a significant reduction in the ability of the cells to proliferate. Furthermore, they exhibited lower infectivity of human peripheral blood mononuclear cells (PBMCs). We also showed that recombinant EBV with deletions of the EBNA3C ORF, as well as a recombinant with residues 621-675 within EBNA3C ORF deleted had diminished abilities to activate CD40. Our study also revealed that the full length (1-992) and 621-675 aa deletions of EBNA3C when co...Continue Reading

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Citations

Feb 11, 2016·Pathogens·Hem Chandra JhaErle S Robertson
Nov 9, 2016·Frontiers in Microbiology·Hem C JhaErle S Robertson

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Methods Mentioned

BETA
transfection
PCR
fluorescence microscopy
confocal microcopy
flow cytometry
nuclear translocation

Software Mentioned

Cell Quest Pro
FlowJo
Odyssey

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