The arenavirus small RING finger Z protein has been shown to be the main driving force of budding for several arenaviruses. This Z budding activity was found to be mediated by the late (L)-domain motifs P(T/S)AP and PPXY, located at the C terminus of Z. Here, we show that the Z protein of Tacaribe virus (TACV), a New World arenavirus, buds efficiently from cells despite lacking the canonical L-domain motifs P(T/S)AP and PPXY. Likewise, potential L-domain motifs ASAP and YLCL present in TACV Z did not exhibit any significant contribution to TACV Z budding activity. Budding of TACV Z was Tsg101 independent but required the activity of Vps4A/B. These results indicate that TACV Z utilizes a budding mechanism distinct from that reported for other arenaviruses.Arenaviruses are enveloped viruses with a bisegmented, negative-strand (NS) RNA genome and a life cycle restricted to the cell cytoplasm (1). Each RNA segment uses an ambisense coding strategy to direct the expression of two genes that are in opposite orientations and separated by a noncoding intergenic region. The large segment (7.2 kb) encodes the late (L) protein, an RNA-dependent RNA polymerase, and the small RING finger protein Z, which is the counterpart of the matrix (M) protein found in many enveloped NS RNA viruses. The small segment (3.5 kb) encodes the viral nucleoprotein (NP) and the glycoprotein precursor