Identification of two types of keratin polypeptides within the acidic cytokeratin subfamily I.
Cytoskeletal filaments of the alpha-keratin type (cytokeratins) are a characteristic of epithelial cells. In diverse mammals (man, cow and rodents) these cytokeratins consist of a family of approximately 20 polypeptides, which may be divided into the more acidic (I) and the more basic (II) subfamilies. These two subfamilies show only limited amino acid sequence homology. In contrast, nucleic acid hybridization experiments and peptide maps have been interpreted to show that polypeptides of the same subfamily share extended sequence homology. We compare two polypeptides of the acidic cytokeratin subfamily, VIb (Mr 54,000) and VII (Mr 50,000), which are co-expressed in large amounts in bovine epidermal keratinocytes. These two epidermal keratins can be distinguished by specific antibodies and show different patterns of expression among several bovine tissues and cultured cells. In addition, they differ in the stability of their complexes with basic keratin polypeptides and in their tryptic peptide maps. The amino acid sequences deduced from the nucleotide sequences of complementary DNA clones containing the 3' ends of the messenger RNAs for these keratins are compared with each other and with available amino acid sequences of human, murine and amphibian epidermal keratins. Bovine keratins VIb and VII share considerable sequence homology in the alpha-helical portion (68% residues identical) but lack significant homology in the extrahelical portion. Bovine keratin VIb shows, in its alpha-helical region, a pronounced sequence homology (88% identity) to the murine epidermal keratin of Mr 59,000. In addition, the non-helical carboxy-terminal regions of both proteins are glycine-rich and contain a canonic sequence GGGSGYGG, which may be repeated several times. Moreover, their mRNAs present a highly conserved stretch of 236 nucleotides containing, in the murine sequence, the end of the coding and all of the non-coding region (81% identical nucleotides). Bovine keratin VII is considerably different from the murine Mr 59,000 keratin but is almost identical to the human cytokeratin number 14 of Mr 50,000, both in the alpha-helical and in the non-alpha-helical regions of the proteins, and the mRNAs of the human and the bovine keratins also display a high homology in their 3' non-coding ends. The results show that in the same species keratins of the same subfamily can differ considerably, whereas equivalent keratin polypeptides of different species are readily identified by characteristic sequence homologies in the alpha-helical and the non-helical regions as well as in the 3' non-coding portions of their mRNAs.(ABSTRACT TRUNCATED AT 400 WORDS)
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003599:Cytoskeleton / Q000032:analysis / Q000648:ultrastructure; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004817:Epidermis / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D007525:Isoelectric Focusing; D007633:Keratins / Q000302:isolation & purification; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000302:isolation & purification; D012333:RNA, Messenger
D010455:Peptides; D012333:RNA, Messenger; D007633:Keratins; D004247:DNA