EBNA2-EBF1 complexes promote MYC expression and metabolic processes driving S-phase progression of Epstein-Barr virus-infected B cells

Epstein-Barr virus (EBV) is a human tumor virus which preferentially infects resting human B cells. Upon infection in vitro, EBV activates and immortalizes these cells. The viral latent protein EBV nuclear antigen 2 (EBNA2) is essential for B cell activation and immortalization; it targets and binds the cellular and ubiquitously expressed DNA-binding protein CBF1, thereby transactivating a plethora of viral and cellular genes. In addition, EBNA2 uses its N-terminal dimerization (END) domain to bind early B cell factor 1 (EBF1), a pioneer transcription factor specifying the B cell lineage. We found that EBNA2 exploits EBF1 to support key metabolic processes and to foster cell cycle progression of infected B cells in their first cell cycles upon activation. The alpha 1-helix within the END domain was found to promote EBF1 binding. EBV mutants lacking the alpha 1-helix in EBNA2 can infect and activate B cells efficiently, but activated cells fail to complete the early S phase of their initial cell cycle. Expression of MYC, target genes of MYC and E2F, as well as multiple metabolic processes linked to cell cycle progression are impaired in EBV Delta alpha 1-infected B cells. Our findings indicate that EBF1 controls B cell activation via EBNA2 and, thus, has a critical role in regulating the cell cycle of EBV-infected B cells. This is a function of EBF1 going beyond its well-known contribution to B cell lineage specification.

Automated summary

EBV is a human tumor virus that preferentially infects resting human B cells. The viral protein EBNA2 plays a crucial role in B cell activation and immortalization by targeting and binding cellular DNA-binding protein CBF1. Our study found that EBNA2 also uses its N-terminal dimerization domain to bind pioneer transcription factor EBF1, supporting metabolic processes and cell cycle progression in infected B cells. These findings indicate the critical role of EBF1 in regulating the cell cycle of EBV-infected B cells.