cDNA cloning and characterization of sciellin, a LIM domain protein of the keratinocyte cornified envelope

Sciellin is a precursor of the cornified envelopes of mammalian keratinizing tissues. We have cloned the cDNA encoding sciellin by screening a human keratinocyte expression library with a sciellin-specific monoclonal antibody. The composite cDNA of 2.35 kilobase pairs encodes a protein of 75.3 kDa with a pi of 10.09, The translated sequence has a central domain containing 16 repeats of 20 amino acids each that is rich in Gin and Lys residues, which are potential transglutaminase substrates, and a carboxyl domain, which contains a single LIM motif, Sciellin cDNA probes hybridize to bands of 3.4 and 4.4 kilobase pairs on Northern blots of cultured human keratinocyte RNA. The gene was mapped to human chromosome band 13q22 by fluorescence in situ hybridization, Radiation hybrid mapping demonstrated that sciellin is linked to the sequence tagged site marker WI-457 with a logarithm of the odds score of 7.77, In situ hybridization of human foreskin tissue sections demonstrated that sciellin is expressed in the stratum granulosum, Immunofluorescent staining with a polyclonal rabbit antibody made to a recombinant sciellin protein showed peripheral cytoplasmic localization in the upper cell. layers of epidermis and in stratified squamous epithelia such as the oral cavity, esophagus, and vagina. Simple and columnar epithelia, with the exception of the amnion, showed no reaction.