Non-protein-coding RNAs (ncRNAs) have recently emerged on the scene of genomic research as prominent players in the regulation of gene expression. Many functionally characterized ncRNAs have been shown to be differentially expressed in various organisms during specific environmental or developmental conditions, thus establishing regulatory networks crucial for shaping cellular life. Here, we show that the expression of vault RNAs (vtRNAs) is specifically up-regulated in human lymphocytes upon infection by gamma-herpesviruses, such as the Epstein-Barr virus and Kaposi's sarcoma virus. vtRNAs are ncRNAs that are integral to the vault complex, a gigantic (13 MDa) hollow ribonucleoprotein particle with a thus far elusive biological role. Stimulation of vtRNA expression by the Epstein-Barr virus was evident for all three canonical vtRNAs (hvg1-hvg3) and also for a novel ncRNA candidate, initially termed CBL-3. This ncRNA shares clear primary- and secondary-structure similarities with the three known vtRNAs. Importantly, CBL-3 co-sediments with intact vault particles in density gradients of various human cell lines, thus strongly indicating this ncRNA as a novel, fourth vault-complex-associated RNA.