Up-regulation of Murid herpesvirus 4 ORF50 by hypoxia: Possible implication for virus reactivation from latency

Murid herpesvirus 4 (MuHV-4) is a member of the Gammaherpesvirus subfamily capable to establish a long-lasting latency and induce occasional malignancies. Because MuHV-4 is associated with cancer in a subset of virus-infected mice and because tumor development is often linked with hypoxia, we studied the influence of hypoxia on the biology of this virus. Using immunofluorescence and FACS analysis we detected increased proportion of MuHV-4 positive cells in the latently infected NB-78 cell line exposed to low oxygen conditions compared to normoxic controls. Moreover, the expression of ORF50, a crucial gene responsible for switch from latency to lytic virus replication, was induced upon the exposure of NB-78 cells to hypoxia. Luciferase reporter assays with ORF50 promoter confirmed the hypoxia-dependent induction. Transient co-transfections with hypoxia inducible factors showed that HIF-2α is a more potent activator of ORF50 expression than HIF-1α. Our results confirm that the MuHV-4 life cycle is influenced by low oxygen concentration.