Cytokeratin expression in simple epithelia. I. Identification of mRNA coding for human cytokeratin no. 18 by a cDNA clone.
To study the regulation of the expression of cytokeratins characteristic of simple epithelia, i.e., human cytokeratins nos. 7, 8, 18, and 19, we prepared several cDNA clones coding for these proteins and their bovine counterparts. In the present study, we describe a cDNA clone of the mRNA coding for human cytokeratin no. 18, which was isolated from an expression library using the monoclonal antibody, KG 8.13. This clone (756 nucleotides, excluding the polyA portion), encodes approximately one-half of the mRNA (approximately 1.4 kb), identifies one mRNA band in Northern-hybridization blots, and specifically selects one mRNA species coding for cytokeratin no. 18, as demonstrated by translation in vitro. Comparison of the deduced amino acid sequence--confirmed by direct amino-acid-sequence analyses of some polypeptide fragments produced by cleavage with cyanogen bromide--indicated that cytokeratin no. 18 is a member of the acidic (type I) subfamily of cytokeratins. It has only limited sequence homologies in common with other intermediate-sized filament proteins, and these are essentially restricted to certain domains of the alpha-helical rod portion. The carboxyterminal tail sequence does not contain glycine-rich elements, thus distinguishing this cytokeratin from those acidic (type I) cytokeratins that are characterized by this feature. The similarities and differences between cytokeratin no. 18 and previously described epidermal cytokeratins are discussed in relation to the differences in the stability of the complexes which this cytokeratin forms with basic (type II) cytokeratins, as well as in relation to possible functional differences of cytokeratins in simple and stratified epithelia.
V
V
Romano
M
M
Hatzfeld
T M
TM
Magin
R
R
Zimbelmann
W W
WW
Franke
G
G
Maier
H
H
Ponstingl
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000235:genetics* / Q000302:isolation & purification; D008099:Liver / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010446:Peptide Fragments / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004274:DNA, Recombinant; D010446:Peptide Fragments; D012333:RNA, Messenger; D007633:Keratins; D004247:DNA