A unique type I keratin intermediate filament group, comprising three highly related proteins and expressed in the inner root sheath of hair follicles, has been identified in both sheep and human. The first members from these species are named oIRSa1 and hIRSa1 and each encodes a protein of 450 amino acids, with compositional characteristics intermediate between those of previously described hair keratin and epidermal cytokeratin type I intermediate filaments. Detection of abundant mRNA transcripts derived from the sheep and human genes by cRNA in situ hybridization only in the inner root sheath and not in the medulla concurs with the findings of earlier ultrastructural analyses that have reported intermediate filaments only in the inner root sheath. Clustering of the IRSa keratin genes is apparent in the genomes of both species. The three hIRSa genes, known to reside on human chromosome 17, are closely linked to three further type I keratin intermediate filament genes of unknown function. This new gene complex, contained almost entirely within a 156 kb BAC (hRPK.142_H_19), is likely to lie near the type I intermediate filament cytokeratin and hair keratin gene loci at 17q12-q21. A phylogenetic analysis including all known human type I intermediate filament cytokeratins, hHa keratins, hIRSa, and hIRSa-linked keratins suggests that origin of the IRSa keratin intermediate filament linkage group preceded origin of most of the epidermal cytokeratins and all hair keratins during emergence of the keratin intermediate filament genes.