Type II keratin cDNAs from the rainbow trout: implications for keratin evolution.
From a teleost fish, the rainbow trout Oncorhynchus mykiss, we have cloned and sequenced cDNAs encoding five different type II keratins. The corresponding protein spots, as separated by 2D-PAGE of trout cytoskeletal preparations, have been identified by peptide mass mapping using MALDI mass spectrometry. Three of the sequenced keratins are expressed in the epidermis (subtype IIe), and two in simple epithelia and mesenchymal cells (subtype IIs). The IIs keratins are both orthologs of human K8. This leaves unsequenced only the trace component S3 of the biochemically established trout keratin catalog. A phylogenetic tree has been constructed from a multiple alignment of the rod domains of the new keratin sequences together with type II sequences from other vertebrates such as shark, zebrafish, and human; lamprey K8 (recently sequenced in our laboratory) has been used as outgroup. This tree suggests, in a highly bootstrap-supported manner, that the teleost IIe keratins diversified independently from the mammalian IIe keratins. In contrast, all the species investigated express K8-like keratins, suggesting that the different IIe branches evolved from K8-like progenitors. The tree also indicates that the published zebrafish sequences represent IIe keratins and that the biochemically identified K8 ortholog in zebrafish has not yet been sequenced.
Institute of Zoology, Johannes Gutenberg University, 55099 Mainz, Germany.
Michael
M
Schaffeld
Mark
M
Haberkamp
Erik
E
Braziulis
Bernhard
B
Lieb
Jürgen
J
Markl
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004817:Epidermis / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D019143:Evolution, Molecular; D006801:Humans; D007633:Keratins / Q000145:classification / Q000235:genetics* / Q000276:immunology; D007798:Lampreys / Q000235:genetics; D008322:Mammals / Q000235:genetics; D008648:Mesoderm / Q000378:metabolism; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics*; D009928:Organ Specificity; D010449:Peptide Mapping; D010802:Phylogeny; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012754:Sharks / Q000235:genetics; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015027:Zebrafish / Q000235:genetics
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D020033:Protein Isoforms; D007633:Keratins