Isolation, sequence and expression of the gene encoding human keratin 13.
Keratins are a family of highly homologous proteins expressed as pairs of acidic and basic forms which make intermediate filaments in epithelial cells. Keratin 13 (K13) is the major acidic keratin, which together with K4, its basic partner, is expressed in the suprabasal layers of non-cornified stratified epithelia. The mechanism which allows mucosal-specific expression of this keratin remains unknown. To provide insight into the tissue-specific expression, we have isolated the human K13 gene by screening a chromosome 17 library with a specific K13 cRNA probe. Sequence analysis of unidirectional deletions produced by transposon Tn3 has revealed that the gene is 4601 nucleotides long and contains seven introns and eight exons. When driven by the CMV promoter, the gene produced K13 protein in MCF-7 cells, which normally do not express this protein. Two transcription-start sites were identified, the major being at 61 and the minor at 63 nucleotides upstream of ATG. The upstream sequence contained a TATA box and several other putative transcription factor binding sites. A single copy of the K13 gene was detected in the human genome by Southern hybridisation and polymerase chain reaction. K13 mRNA shows differential expression in cultured keratinocytes, and in A431 cells the RNA levels remained independent of calcium concentrations in the culture medium. Characterisation of the human K13 gene will facilitate elucidation of the molecular mechanism regulating K13 expression in mucosal tissues.
Waseem
A
A
Department of Craniofacial Development, Guy's Dental School, Floor 28, Guy's Tower, London Bridge, London, SE1 9RT, UK. a.wasseem@umds.ac.uk
Alam
Y
Y
Dogan
B
B
White
K N
KN
Leigh
I M
IM
Waseem
N H
NH
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D001943:Breast Neoplasms; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17; D017931:DNA Primers; D005091:Exons; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D007624:KB Cells; D015603:Keratinocytes / Q000378:metabolism*; D007633:Keratins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014157:Transcription Factors; D007633:Keratins