We present the initial characterization of a novel family of genes that rearrange in T cells, but do not encode either of the defined (alpha/beta) subunits of the clone-specific heterodimer of the T cell receptor. The family comprises at least three variable (V) gene segments, three constant (C) gene segments, and three junction (J) gene segments. In a cloned cytolytic T lymphocyte, 2C, one of each of these fragments has productively rearranged to yield an expressed VJC transcription unit, which shows no evidence for somatic mutation. Short sequences similar to those implicated in immunoglobulin gene and T cell receptor beta chain gene rearrangement flank the V and J segments of this family. The linkage of two of the three V gene segments has been determined: the segments lie approximately 2.5 kb apart, and are arranged head-to-head. The inverted arrangement may cast light upon the mechanisms utilized by lymphocytes for gene rearrangement.