The ciliated protozoan Tetrahymena thermophila undergoes extensive programmed DNA rearrangements during the development of a somatic macronucleus from the germ line micronucleus in its sexual cycle. To investigate the relationship between programmed DNA rearrangements and transposable elements, we identified several members of a family of non-long terminal repeat (LTR) retrotransposons (retroposons) in T. thermophila, the first characterized in the ciliated protozoa. This multiple-copy retrotransposon family is restricted to the micronucleus of T. thermophila. The REP (Tetrahymena non-LTR retroposon) elements encode an ORF2 typical of non-LTR elements that contains apurinic/apyrimidinic endonuclease (APE) and reverse transcriptase (RT) domains. Phylogenetic analysis of the RT and APE domains indicates that the element forms a deep-branching clade within the non-LTR retrotransposon family. Northern analysis with a probe to the conserved RT domain indicates that transcripts from the element are small and heterogeneous in length during early macronuclear development. The presence of a repeated transposable element in the genome is consistent with the model that programmed DNA deletion in T. thermophila evolved as a method of eliminating deleterious transposons from the somatic macronucleus.Developmentally programmed DNA rearrangements occur in a wide variety of organisms (reviewed in reference 5). Functions such as altering gene dosage or directly regulating gene expression have been assigned to many but not all examples of programmed DNA rearrangements. A clinically important example of a programmed DNA rearrangement is V(D)J recombination (2). In addition, a variety of mammalian parasites use programmed DNA rearrangements to vary their surface antigens to avoid host immune response (4). The function of other programmed DNA rearrangements is not as clear. The extensive genome rearrangements that occur during nuclear development in the ciliated protozoa provide an example of programmed DNA rearrangements with poorly understood function.Like all ciliated protozoa, the oligohymenopheran Tetrahymena thermophila displays nuclear dimorphism with a mostly transcriptionally silent diploid germ line nucleus (micronucleus) and a polyploid and transcriptionally active somatic nucleus (macronucleus) within the same cell. The macronucleus develops from a mitotic product of the micronucleus during conjugation. When two cells of different mating types conjugate, the micronucleus in each divides meiotically and mitotically to generate a haploid gametic nucleus that is reciprocally exchanged and fuses with that of its partner to form a zygotic nucleus. This zygotic nucleus divides and from one of the products develops a new macronucleus, while the old macronucleus is concurrently degraded. In T. thermophila, macronuclear development involves extensive programmed DNA rearrangements, including chromosome fragmentation, DNA amplification, and site-specific interstitial DNA deletion (12). Interstitial DNA deletion is responsible for t...