A protein (ORF2) encoded by the latency related gene of bovine herpesvirus 1 interacts with DNA. 2 (BMPR2) was intriguing because it encodes a serine/threonine receptor kinase that promotes neuronal differentiation and inhibits cell death. Another differentially expressed gene encodes a protein kinase (Akt3), which is usually significant because Akt activity enhances cell survival and is linked to herpes simplex virus 1 latency and neuronal survival. Additional studies exhibited ORF2 increased Akt3 steady-state protein levels and interacted with Akt3 in transfected Neuro-2A cells, which correlated with Akt3 activation. Conversely, expression of Wnt antagonists increased during reactivation from latency. Collectively, these studies suggest Wnt signaling cooperates with LR gene products, in particular ORF2, to promote latency. IMPORTANCE Lifelong BoHV-1 latency primarily occurs in sensory neurons. The synthetic corticosteroid dexamethasone consistently induces reactivation from latency in calves. RNA sequencing studies revealed 102 genes associated with the Wnt/-catenin signaling pathway are differentially regulated during the latency-reactivation cycle. Two protein kinases associated with the Wnt pathway, Akt3 and BMPR2, were expressed at higher levels Isatoribine monohydrate during latency but were repressed during reactivation. Furthermore, five genes encoding soluble Wnt antagonists and -catenin-dependent transcription Isatoribine monohydrate inhibitors were induced during reactivation from latency. These findings are important because Wnt, BMPR2, and Akt3 promote neurogenesis and cell survival, processes crucial for lifelong viral latency. In transfected neuroblastoma cells, a viral protein expressed during latency (ORF2) interacts with and enhances Akt3 protein kinase activity. These findings provide insight into how cellular factors associated with the Wnt signaling pathway cooperate with LR gene products to regulate the BoHV-1 latency-reactivation cycle. = 1.54e?3; 30 min after DEX, = 7.16e?6; 1.5 h after DEX, = 3.55e?3; and 3 h after DEX, = 8.15e?4. Ingenuity pathway analysis (IPA) identified 102 unique genes or gene families involved with the Wnt/-catenin signaling pathway that were differentially regulated across these Isatoribine monohydrate Isatoribine monohydrate time points: 29 in uninfected TG, 72 at 30 min after DEX, 35 at 1.5 h after DEX, and 58 at 3 h after DEX (Fig. 1). Table 1 provides a summary of the statistical analysis of the read data. Open in a separate windows FIG 1 Heat map summarizing differentially expressed genes that are associated Dnm2 with the Wnt/-catenin signaling pathway in TG during BoHV-1 latency-reactivation cycle. RNA sequencing was performed using TG from calves for each group (one uninfected, three latently infected, and eight after DEX treatment to induce reactivation [two calves at 30 min, three calves at 1.5 h, and three calves at 3 h after DEX treatment]). Genes associated with the Wnt/-catenin signaling pathway in TG of uninfected calves were compared to those of calves latently infected. Genes associated with the Wnt/-catenin signaling pathway in TG during early says of reactivation (30 min, 1.5 h, and 3 h after DEX treatment) were compared to those in latency. *, fold change in latently infected TG compared to uninfected TG; **, fold change after DEX treatment compared to latently infected TG. Statistical analyses of the read data are summarized in Table 1. TABLE 1 Summary of RNA sequencing of TG from calves used in this study 0.05) in the numbers of GNAQ+ neurons as determined by a Student test. Magnification is approximately 200. Similarly, we examined BMPR2 expression in TG neurons. BMPR2 was detected in a small subset of TG neurons of uninfected calves (16%); however, the number of BMPR2+ neurons increased significantly in latently infected tissue (75%) (Fig. 4A and.