Characterization of human cytokeratin 2, an epidermal cytoskeletal protein synthesized late during differentiation.
Among the more than 30 different human proteins of the cytokeratin (CK) group of intermediate filament (IF) proteins, the significance of the epidermal polypeptide CK 2 (Moll et al., 1982, Cell 31, 11-24) has been repeatedly questioned in the literature. Here, we show, by in vitro translation and protein gel electrophoresis, that human epidermis from various body sites does indeed contain relatively large amounts of mRNA encoding a distinct polypeptide comigrating with native epidermal CK 2. We also report the isolation of a cDNA clone encoding the complete sequence of CK 2, which is a type II CK different from--but related to--epidermal CKs 1 and 5 on the one hand and corneal CK 3 on the other. The mRNA of approximately 2.6 kb encodes a polypeptide of 645 amino acids and M(r) 65,852, in good agreement with the value of 65.5 kDa previously estimated from gel electrophoresis. This human CK, the largest so far known, displays several features typical of CKs of stratified epithelia, including numerous repeats of glycine-rich tetrapeptides in the head and tail domains. Northern blot and in situ hybridizations have shown that CK 2 is expressed strictly suprabasally, usually starting in the third or fourth cell layer of epidermis, and this was confirmed at the protein level by immunohistochemistry using CK 2-specific antibodies. The protein has been detected as a regular epidermal component in skin samples from different body sites, albeit as a minor CK in "soft skin" (e.g., breast nipple, penile shaft, axilla), but not in foreskin epithelium and in other epithelia, in squamous metaplasias and carcinomas, or in cultured cell lines derived therefrom. We propose that CK 2 is a late cytoskeletal IF addition synthesized during maturation of epidermal keratinocytes which probably contributes to terminal cornification.
Collin
C
C
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Moll
R
R
Kubicka
S
S
Ouhayoun
J P
JP
Franke
W W
WW
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D015139:Blotting, Southern; D002454:Cell Differentiation; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D000078404:Epidermal Cells; D004817:Epidermis / Q000502:physiology*; D005260:Female; D006801:Humans; D007633:Keratins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008297:Male; D008969:Molecular Sequence Data; D009061:Mouth Mucosa / Q000166:cytology / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D012867:Skin / Q000166:cytology / Q000378:metabolism*
D000906:Antibodies; D009838:Oligodeoxyribonucleotides; D012313:RNA; D007633:Keratins; D004247:DNA