Correlations between the T cell receptor (TcR) V gene usage and the specificity of T cells have been primarily described for major histocompatibility complex (MHC) class II-restricted helper T cell responses. In the present study the TcR genes expressed by MHC class I-restricted murine cytotoxic T cells (CTL) specific for a major epitope of the lymphocytic choriomeningitis virus (LCMV), LCMV-GP2(275-289), were investigated. The TcR primary structure of an LCMV-GP2(275-289) specific H-2Db-restricted CTL clone has been determined. It uses a member of the V alpha 4 family joined to J alpha AN14.4 for the alpha chain and V beta 10 rearranged to D beta 2.1 and J beta 2.4 for its beta chain. Four other independent LCMV-GP2(275-289) specific H-2Db-restricted CTL clones also expressed V alpha 4 and V beta 10 gene elements. Furthermore, V alpha 4 and V beta 10 were preferentially expressed by polyclonal CTL of C57BL/6 origin specific for LCMV. These results suggest that both TcR V alpha and V beta regions are important for the recognition of the LCMV-GP2(275-289) epitope on H-2Db molecules.