Full-length cDNA sequences of two class I major histocompatibility complex molecules from the DA strain of Rattus norvegicus are reported. One codes for the classical class I restriction element RTL.Aa, which maps to the locus in the rat major histocompatibility complex homologous to H-2K in the mouse. The other probably codes for a soluble nonclassical class I molecule present in DA rat serum; a short deletion in the fifth exon implies that the translated product will terminate in the membrane-spanning region. These sequences have been compared with mouse classical class I sequences as well as with three published rat class I cDNA partial sequences. The results show, first, that "locus-specific" substitutions from the H-2K, H-2D, and H-2L data set are scrambled in the RTl.A molecule; a majority of these substitutions have H-2D/L-specific features. Second, the data show that the four rat sequences are strikingly similar to one another regardless of locus or haplotype of origin; they share a number of apparently species-specific features that distinguish them all from mouse classical class I sequences, which likewise share distinctive features of their own. The results suggest that segmental sequence exchange plays a major role in determining the evolution of sequence in class I masjor histocompatibility complex molecules.Some multigene families display a degree of species specificity at the level of DNA sequence, as if the whole family were evolving within a species as a unit (1-3). Such concerted evolution has been suspected in mouse major histocompatibility complex (MHC) class I genes because of the lack of clear locus-specific sequence variation distinguishing the widely separated H-2K and H-2D end classical loci (4, 5). Short segmental exchanges of sequence are indeed occurring between widely separated class I MHC loci in laboratory mice (6). Nevertheless it remains controversial whether such processes contribute significantly to the pattern of evolution of class I genes (7), since sequences of class I genes from a closely related murine species such as the rat are not yet available. Rattus norvegicus is in the same subfamily, the Murinae, as the mouse. The rat MHC, RTI, is like H-2 in structure, with a classical class I gene, RTJ.A, at the left end of the MHC in the same location as H-2K and a long array of other class I genes to the right (8). In the middle is a region specifying class II genes that closely resembles the homologous region of H-2 (9). It is most likely that the translocation of class I genes that formed the H-2K end was a unique event occurring shortly before the separation of the rat and mouse lineages (7). Rat class I gene sequences thus provide a stringent test of the concerted evolution hypothesis. With simple divergent evolution alone, the sequence of an RTI.A allele should have greater similarity to H-2K on average than to other rat class I sequences, since RTI.A and H-2K share a more recent common ancestor than either does with genes at the other end of the MHC. If, on the ot...