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. 1986 Jan 24;14(2):751–763. doi: 10.1093/nar/14.2.751

Nonepidermal members of the keratin multigene family: cDNA sequences and in situ localization of the mRNAs.

B Knapp, M Rentrop, J Schweizer, H Winter
PMCID: PMC339462  PMID: 2418416

Abstract

A keratin set which consists of a type I 47kd and a type II 57kd protein occurs as a major constituent of the keratin patterns of various internal stratified epithelia of the mouse. We have isolated specific cDNA clones of the two complementary keratin subunits from a cDNA library constructed with polyA+RNA of mouse tongue epithelium and present the complete nucleotide and deduced amino acid sequences of the 57kd protein and about 75% of the corresponding data of the 47kd protein. The comparison of the sequence data with those of known epidermal keratin mRNAs coding for the two types of keratin proteins reveals a fundamentally identical and type-specific organization of the mRNAs into both highly conserved and variable domains. In order to avoid cross-reactions with other members of the keratin multigene family, appropriately taylored 35S-labeled cDNA probes comprising the low and non-homologous 3' coding and noncoding domains of the mRNAs were used for in situ hybridization to tissue sections. The localization and distribution of the corresponding transcripts indicates a strongly compartmentalized keratin expression in mouse tongue epithelium.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  2. Baralle F. E., Brownlee G. G. AUG is the only recognisable signal sequence in the 5' non-coding regions of eukaryotic mRNA. Nature. 1978 Jul 6;274(5666):84–87. doi: 10.1038/274084a0. [DOI] [PubMed] [Google Scholar]
  3. Chou P. Y., Fasman G. D. Prediction of beta-turns. Biophys J. 1979 Jun;26(3):367–383. doi: 10.1016/S0006-3495(79)85259-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crewther W. G., Inglis A. S., McKern N. M. Amino acid sequences of alpha-helical segments from S-carboxymethylkerateine-A. Complete sequence of a type-II segment. Biochem J. 1978 Aug 1;173(2):365–371. doi: 10.1042/bj1730365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  6. Franke W. W., Schiller D. L., Hatzfeld M., Winter S. Protein complexes of intermediate-sized filaments: melting of cytokeratin complexes in urea reveals different polypeptide separation characteristics. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7113–7117. doi: 10.1073/pnas.80.23.7113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Franke W. W., Schmid E., Schiller D. L., Winter S., Jarasch E. D., Moll R., Denk H., Jackson B. W., Illmensee K. Differentiation-related patterns of expression of proteins of intermediate-size filaments in tissues and cultured cells. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):431–453. doi: 10.1101/sqb.1982.046.01.041. [DOI] [PubMed] [Google Scholar]
  8. Fuchs E., Green H. Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell. 1981 Sep;25(3):617–625. doi: 10.1016/0092-8674(81)90169-0. [DOI] [PubMed] [Google Scholar]
  9. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  10. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hagenbüchle O., Santer M., Steitz J. A., Mans R. J. Conservation of the primary structure at the 3' end of 18S rRNA from eucaryotic cells. Cell. 1978 Mar;13(3):551–563. doi: 10.1016/0092-8674(78)90328-8. [DOI] [PubMed] [Google Scholar]
  12. Hanukoglu I., Fuchs E. The cDNA sequence of a Type II cytoskeletal keratin reveals constant and variable structural domains among keratins. Cell. 1983 Jul;33(3):915–924. doi: 10.1016/0092-8674(83)90034-x. [DOI] [PubMed] [Google Scholar]
  13. Hanukoglu I., Fuchs E. The cDNA sequence of a human epidermal keratin: divergence of sequence but conservation of structure among intermediate filament proteins. Cell. 1982 Nov;31(1):243–252. doi: 10.1016/0092-8674(82)90424-x. [DOI] [PubMed] [Google Scholar]
  14. Johnson L. D., Idler W. W., Zhou X. M., Roop D. R., Steinert P. M. Structure of a gene for the human epidermal 67-kDa keratin. Proc Natl Acad Sci U S A. 1985 Apr;82(7):1896–1900. doi: 10.1073/pnas.82.7.1896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jorcano J. L., Franz J. K., Franke W. W. Amino acid sequence diversity between bovine epidermal cytokeratin polypeptides of the basic (type II) subfamily as determined from cDNA clones. Differentiation. 1984;28(2):155–163. doi: 10.1111/j.1432-0436.1984.tb00278.x. [DOI] [PubMed] [Google Scholar]
  16. Jorcano J. L., Rieger M., Franz J. K., Schiller D. L., Moll R., Franke W. W. Identification of two types of keratin polypeptides within the acidic cytokeratin subfamily I. J Mol Biol. 1984 Oct 25;179(2):257–281. doi: 10.1016/0022-2836(84)90468-6. [DOI] [PubMed] [Google Scholar]
  17. Krieg T. M., Schafer M. P., Cheng C. K., Filpula D., Flaherty P., Steinert P. M., Roop D. R. Organization of a type I keratin gene. Evidence for evolution of intermediate filaments from a common ancestral gene. J Biol Chem. 1985 May 25;260(10):5867–5870. [PubMed] [Google Scholar]
  18. Lawrence J. B., Singer R. H. Quantitative analysis of in situ hybridization methods for the detection of actin gene expression. Nucleic Acids Res. 1985 Mar 11;13(5):1777–1799. doi: 10.1093/nar/13.5.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marchuk D., McCrohon S., Fuchs E. Complete sequence of a gene encoding a human type I keratin: sequences homologous to enhancer elements in the regulatory region of the gene. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1609–1613. doi: 10.1073/pnas.82.6.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marchuk D., McCrohon S., Fuchs E. Remarkable conservation of structure among intermediate filament genes. Cell. 1984 Dec;39(3 Pt 2):491–498. doi: 10.1016/0092-8674(84)90456-2. [DOI] [PubMed] [Google Scholar]
  21. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  22. Moll R., Franke W. W., Schiller D. L., Geiger B., Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell. 1982 Nov;31(1):11–24. doi: 10.1016/0092-8674(82)90400-7. [DOI] [PubMed] [Google Scholar]
  23. Osterburg G., Glatting K. H., Sommer R. Computer programs for the analysis and the management of DNA sequences. Nucleic Acids Res. 1982 Jan 11;10(1):207–216. doi: 10.1093/nar/10.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Parry D. A., Crewther W. G., Fraser R. D., MacRae T. P. Structure of alpha-keratin: structural implication of the amino acid sequences of the type I and type II chain segments. J Mol Biol. 1977 Jun 25;113(2):449–454. doi: 10.1016/0022-2836(77)90153-x. [DOI] [PubMed] [Google Scholar]
  25. Potten C. S., Schofield R., Lajtha L. G. A comparison of cell replacement in bone marrow, testis and three regions of surface epithelium. Biochim Biophys Acta. 1979 Aug 10;560(2):281–299. doi: 10.1016/0304-419x(79)90022-2. [DOI] [PubMed] [Google Scholar]
  26. Rieger M., Jorcano J. L., Franke W. W. Complete sequence of a bovine type I cytokeratin gene: conserved and variable intron positions in genes of polypeptides of the same cytokeratin subfamily. EMBO J. 1985 Sep;4(9):2261–2267. doi: 10.1002/j.1460-2075.1985.tb03924.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schweizer J., Goerttler K. Synthesis in vitro of keratin polypeptides directed by mRNA isolated from newborn and adult mouse epidermis. Eur J Biochem. 1980 Nov;112(2):243–249. doi: 10.1111/j.1432-1033.1980.tb07200.x. [DOI] [PubMed] [Google Scholar]
  30. Schweizer J., Kinjo M., Fürstenberger G., Winter H. Sequential expression of mRNA-encoded keratin sets in neonatal mouse epidermis: basal cells with properties of terminally differentiating cells. Cell. 1984 May;37(1):159–170. doi: 10.1016/0092-8674(84)90311-8. [DOI] [PubMed] [Google Scholar]
  31. Schweizer J., Winter H. Changes in regional keratin polypeptide patterns during phorbol ester-mediated reversible and permanently sustained hyperplasia of mouse epidermis. Cancer Res. 1982 Apr;42(4):1517–1529. [PubMed] [Google Scholar]
  32. Schweizer J., Winter H., Hill M. W., Mackenzie I. C. The keratin polypeptide patterns in heterotypically recombined epithelia of skin and mucosa of adult mouse. Differentiation. 1984;26(2):144–153. doi: 10.1111/j.1432-0436.1984.tb01388.x. [DOI] [PubMed] [Google Scholar]
  33. Schweizer J., Winter H. Keratin biosynthesis in normal mouse epithelia and in squamous cell carcinomas. mRNA-dependent alterations of the primary structure of distinct keratin subunits in tumors. J Biol Chem. 1983 Nov 10;258(21):13268–13272. [PubMed] [Google Scholar]
  34. Smith H. O., Birnstiel M. L. A simple method for DNA restriction site mapping. Nucleic Acids Res. 1976 Sep;3(9):2387–2398. doi: 10.1093/nar/3.9.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Steinert P. M., Jones J. C., Goldman R. D. Intermediate filaments. J Cell Biol. 1984 Jul;99(1 Pt 2):22s–27s. doi: 10.1083/jcb.99.1.22s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Steinert P. M., Parry D. A., Idler W. W., Johnson L. D., Steven A. C., Roop D. R. Amino acid sequences of mouse and human epidermal type II keratins of Mr 67,000 provide a systematic basis for the structural and functional diversity of the end domains of keratin intermediate filament subunits. J Biol Chem. 1985 Jun 10;260(11):7142–7149. [PubMed] [Google Scholar]
  37. Steinert P. M., Parry D. A., Racoosin E. L., Idler W. W., Steven A. C., Trus B. L., Roop D. R. The complete cDNA and deduced amino acid sequence of a type II mouse epidermal keratin of 60,000 Da: analysis of sequence differences between type I and type II keratins. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5709–5713. doi: 10.1073/pnas.81.18.5709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Steinert P. M., Rice R. H., Roop D. R., Trus B. L., Steven A. C. Complete amino acid sequence of a mouse epidermal keratin subunit and implications for the structure of intermediate filaments. Nature. 1983 Apr 28;302(5911):794–800. doi: 10.1038/302794a0. [DOI] [PubMed] [Google Scholar]
  39. Sun T. T., Eichner R., Nelson W. G., Tseng S. C., Weiss R. A., Jarvinen M., Woodcock-Mitchell J. Keratin classes: molecular markers for different types of epithelial differentiation. J Invest Dermatol. 1983 Jul;81(1 Suppl):109s–115s. doi: 10.1111/1523-1747.ep12540831. [DOI] [PubMed] [Google Scholar]
  40. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tseng S. C., Jarvinen M. J., Nelson W. G., Huang J. W., Woodcock-Mitchell J., Sun T. T. Correlation of specific keratins with different types of epithelial differentiation: monoclonal antibody studies. Cell. 1982 Sep;30(2):361–372. doi: 10.1016/0092-8674(82)90234-3. [DOI] [PubMed] [Google Scholar]
  42. Tyner A. L., Eichman M. J., Fuchs E. The sequence of a type II keratin gene expressed in human skin: conservation of structure among all intermediate filament genes. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4683–4687. doi: 10.1073/pnas.82.14.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wickens M. P., Buell G. N., Schimke R. T. Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid, and ovalbumin mRNAs. Optimization for full length second strand synthesis by Escherichia coli DNA polymerase I. J Biol Chem. 1978 Apr 10;253(7):2483–2495. [PubMed] [Google Scholar]
  44. Wu Y. J., Parker L. M., Binder N. E., Beckett M. A., Sinard J. H., Griffiths C. T., Rheinwald J. G. The mesothelial keratins: a new family of cytoskeletal proteins identified in cultured mesothelial cells and nonkeratinizing epithelia. Cell. 1982 Dec;31(3 Pt 2):693–703. doi: 10.1016/0092-8674(82)90324-5. [DOI] [PubMed] [Google Scholar]

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