Avian keratin genes. II. Chromosomal arrangement and close linkage of three gene families.

We describe the isolation and characterization of a set of overlapping cosmid clones that contain chicken keratin genes. The 100 kb (1 kb = 10(3) base-pairs) of DNA represented in these clones contains a cluster of 18 feather keratin genes spanning 53 kb of DNA. The feather keratin genes are spaced about 3 kb apart and at least 11 of them have the same transcriptional orientation. Southern analysis using oligonucleotide probes made from highly conserved portions of the 5' non-coding, intron and 3' non-coding regions, respectively, indicate that these sequences have been highly conserved among the gene family as a whole, with only one or two exceptions in each case. The presence of some regularly repeated restriction enzyme sites are indicative of tandem duplication events in the recent history of the feather keratin gene family. The feather keratin gene locus is flanked on both sides by related types of keratin genes. On the 5' side of the feather gene cluster are three keratin genes (designated feather-like) that are located 5 kb from the last feather keratin gene and are spaced about 4 kb apart. On the 3' side of the feather gene cluster, 21 kb from the last feather keratin gene, lies a cluster of four genes that encode claw keratins. These genes are spaced about 1 kb apart and appear to be divergently transcribed. Partial DNA sequence analysis of the feather-like gene lying proximal to the feather keratin gene cluster demonstrated that it encodes a protein of 115 amino acid residues that is 80% homologous to the feather keratins at both the DNA and amino acid sequence levels. The feather-like gene(s) are expressed in both embryonic and adult (post-hatch) chick feathers and at a very low level in embryonic scale tissue. These genes therefore form a new family of feather proteins that is distinct from the previously characterized feather keratins.