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Cloning and characterization of keratin D, a murine endodermal cytoskeletal protein induced during in vitro differentiation of F9 teratocarcinoma cells

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Summary

We have identified a cDNA coding for the murine keratin D from a collection of clones representing F9 teratocarcinoma stem cell mRNA sequences. These sequences are synthesized specifically after the addition of retinoic acid and cAMP to the culture medium. The clone is 1,382 nucleotides long and contains the entire information for the active polypeptide, the complete 3′ end and most, if not all, of the 5′ non-coding region. The mRNA is found in hepatocytes, in PYS-2 cells (an endodermal cell line) and in differentiated (retinoic-acid-treated) F9 cells, but not in untreated F9 cells. The length of the mRNA is 1.4 kb, as estimated by Northern blot hybridization. Southern hybridization performed under very stringent conditions detects a single fragment hybridizing strongly with the cloned cDNA, suggesting that the mouse genome contains only one or very few copies of this gene. We present the first complete sequence of a keratin expressed in simple epithelia, i.e. keratin D, and discuss its structural features.

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References

  • Duprey P, Morello D, Vasseur M, Babinet C, Condamine H, Brulet P, Jacob F (1985) Expression of the cytokeratin endo A gene during early mouse embryogenesis. Proc Natl Acad Sci (USA) 82:8535–8539

    Google Scholar 

  • Franke WW, Denk H, Kalt R, Schmid E (1981) Biochemical and immunological identification of cytokeratin proteins present in hepatocytes of mammalian liver tissue. Exp Cell Res 131:299–318

    Google Scholar 

  • Garnier J, Osguthorpe DJ, Robson B (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120

    Google Scholar 

  • Hogan BLM, Barlow DP, Tilly R (1983) F9 teratocarcinoma cells as a model system for the differentiation of parietal endoderm and visceral endoderm in the mouse embryo. Cancer Surv 2:115–140

    Google Scholar 

  • Huynh TV, Young RA, Davis RW (1985) Construction and screening cDNA libraries in gt10 and gt11. In: Glover DM (ed) DNA cloning. VI. A practical approach. IRL Press, Oxford Washington, pp 49–78

    Google Scholar 

  • Jackson BW, Grund C, Schmid E, Bürki K, Franke WW, Illmensee K (1980) Formation of cytoskeletal elements during mouse embryogenesis. Intermediate filaments of the cytokeratin type and desmosomes in preimplantation embryos. Differentiation 17:161–179

    Google Scholar 

  • Jackson BW, Grund C, Winter S, Franke WW, Illmensee K (1981) Formation of cytoskeletal elements during mouse embryogenesis. II. Epithelial differentiation and intermediate-sized filaments in early postimplantation embryos. Differentiation 20:203–216

    Google Scholar 

  • Jorcano JL, Rieger M, Franz JK, Schiller DL, Moll R, Franke WW (1984a) Identification of two types of keratin polypeptides within the acidic cytokeratin subfamily I. J Mol Biol 179:257–281

    Google Scholar 

  • Jorcano JL, Franz JK Franke WW (1984b) Amino acid sequence diversity between bovine epidermal cytokeratin polypeptides of the basic (type II) subfamily as determined from cDNA clones. Differentiation 28:155–163

    Google Scholar 

  • Kemler R, Brûlet P, Schnebelen MT, Gaillard J, Jacob F (1981) Reactivity of monoclonal antibodies against intermediate filament proteins during embryonic development. J Embryol Exp Morphol 64:45–60

    Google Scholar 

  • Kurkinen M, Cooper AR, Barlow DP, Jenkins JR, Hogan BLM (1983) Gene expression during parietal endoderm differentiation in mouse embryos and teratocarcinoma cells. In: Silver LM Martin GR, Strickland S (eds) Teratocarcinoma stem cells. Cold Spring Harbor Laboratory, New York, pp 389–402

    Google Scholar 

  • Levine RA, LaRosa JG, Gudas LJ (1984) Isolation of cDNA clones for genes exhibiting reduced expression after differentiation of murine teratocarcinoma stem cell. Mol Cell Biol 4:2142–2150

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base-specific chemical cleavages. Meth Enzymol 65:499–559

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Moll R, Franke WW, Schiller DL, Geiger B, Krepler R (1982) The catalog of human cytokeratin polypeptides: patterns of expression of specific cytokeratins in normal epithelia, tumors, and cultured cells. Cell 31:11–24

    Google Scholar 

  • Osborn M, Weber K (1983) Biology of disease. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest 48:372–394

    Google Scholar 

  • Oshima RG, Howe WE, Klier FG, Adamson ED, Shevinsky LH (1983) Intermediate filament protein synthesis in preimplantation murine embryos. Dev Biol 99:447–455

    Google Scholar 

  • Oshima RG, Howe WE, Tabor JM, Trevor K (1983) Cytoskeletal proteins as markers for embryonal carcinoma differentiation. In: Silver LM, Martin GR, Strickland S (eds) Teratocarcinoma stem cells. Cold Spring Harbor Laboratory, New York, pp 51–62

    Google Scholar 

  • Ramaekers F, Shaap H, Mulder M, Huysmans A, Vooijs P (1984) Cytokeratin filament expression during in vitro teratocarcinoma cell differentiation as detected by a monoclonal antibody. Cell Biol Int Rep 8:507–518

    Google Scholar 

  • Romano V, Hatzfeld M, Magin TM, Zimbelmann R, Franke WW, Meier G, Postingl H (1986) Cytokeratin expression in simple epithelia. I. Identification of mRNA coding for human cytokeratin no. 18 by a cDNA clone. Differentiation 30:244–253

    Google Scholar 

  • Singer PA, Trevor K, Oshima R (1986) Molecular cloning and characterization of the Endo B cytokeratin expressed in preimplantation mouse embryos. J Biol Chem 261:538–547

    Google Scholar 

  • Strickland S, Mahdavi V (1978) The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell 15:393–404

    Google Scholar 

  • Strickland S, Smith KK, Marotti KR (1980) Hormonal induction of differentiation in teratocarcinoma stem cells: generation of parietal endoderm by retinoic acid and dibutyryl cAMP. Cell 21:347–355

    Google Scholar 

  • Sun TT, Eichner R, Schermer A, Cooper D, Nelson WG, Weiss RA (1984) Classification, expression, and possible mechanisms of evolution of mammalian epithelial keratins: a unifying model. In: Levine AT, Vande Woude GF, Topp WC, Watson JT (eds) Cancer cells I. The transformed phenotype. Cold Spring Harbor Laboratory, New York, pp 169–176

    Google Scholar 

  • Vasseur M, Duprey P, Marle C, Brulet P, Jacob F (1985) One gene and one pseudogene for the cytokeratin Endo A. Proc Natl Acad Sci (USA) 82:1155–1159

    Google Scholar 

  • Wang SA, LaRosa GJ (1985) Molecular cloning of gene sequences transcriptionally regulated by retinoic acid and dibutyryl cyclic AMP in cultured mouse teratocarcinoma cells. Dev Biol 107:75–86

    Google Scholar 

  • Weber K, Geisler N (1984) Intermediate filaments — from wool α-keratins to neurofilaments: A structural overview. In: Levine AJ, Vande Woude GF, Topp WC, Watson JD (eds) Cancer cells I. The transformed phenotype. Cold Spring Harbor Laboratory, New York, pp 153–159

    Google Scholar 

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Alonso, A., Weber, T. & Jorcano, J.L. Cloning and characterization of keratin D, a murine endodermal cytoskeletal protein induced during in vitro differentiation of F9 teratocarcinoma cells. Roux's Arch Dev Biol 196, 16–21 (1987). https://doi.org/10.1007/BF00376018

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