Interferon (IFN) type I induces the expression of antiviral proteins such as 2',5'-oligoadenylate synthetases (OAS). The enzyme OAS is activated by dsRNA to produce 5'-phosphorylated, 2-5-linked oligoadenylates (2-5A) that activate RNaseL which, in turn, triggers RNA breakdown, leading to multiple biological functions. Although RNaseL is required for IFN antiviral function, there are many aspects of the molecular mechanisms that remain obscure. Here, we have used microarray analyses from human HeLa cells infected with vaccinia virus (VACV) recombinants expressing OAS-RNaseL enzymes (referred as 2-5A system) with the aim to identify host genes that are up- or down-regulated in the course of infection by the activation of this antiviral pathway. We found that activation of the 2-5A system from VACV recombinants produces a remarkable stimulation of transcription for genes that regulate many cellular processes, like those that promote cell growth arrest, GADD45B and KCTD11, apoptosis as CUL2, PDCD6, and TNFAIP8L2, IFN-stimulated genes as IFI6, and related to tumor suppression as PLA2G2A. The 2-5A system activation produces down-regulation of transcription of some genes that promote cell growth as RUNX2 and ESR2 and of genes in charge to maintain mitochondria homeostasis as MIPEP and COX5A. These results reveal new genes induced in response to the activation of the 2-5A system with roles in apoptosis, translational control, cell growth arrest, and tumor suppression.