Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1990 Dec 1;111(6):2587–2600. doi: 10.1083/jcb.111.6.2587

Structure and expression of genes for a class of cysteine-rich proteins of the cuticle layers of differentiating wool and hair follicles

PMCID: PMC2116416  PMID: 1703541

Abstract

The major histological components of the hair follicle are the hair cortex and cuticle. The hair cuticle cells encase and protect the cortex and undergo a different developmental program to that of the cortex. We report the molecular characterization of a set of evolutionarily conserved hair genes which are transcribed in the hair cuticle late in follicle development. Two genes were isolated and characterized, one expressed in the human follicle and one in the sheep follicle. Each gene encodes a small protein of 16 kD, containing greater than 50 cysteine residues, ranging from 31 to 36 mol% cysteine. Their high cysteine content and in vitro expression data identify them as ultra-high-sulfur (UHS) keratin proteins. The predicted proteins are composed almost entirely of cysteine-rich and glycine-rich repeats. Genomic blots reveal that the UHS keratin proteins are encoded by related multigene families in both the human and sheep genomes. Tissue in situ hybridization demonstrates that the expression of both genes is localized to the hair fiber cuticle and occurs at a late stage in fiber morphogenesis.

Full Text

The Full Text of this article is available as a PDF (4.1 MB).

Selected References

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

  1. Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  5. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  6. Choo K. H., Filby G., Greco S., Lau Y. F., Kan Y. W. Cosmid vectors for high efficiency DNA-mediated transformation and gene amplification in mammalian cells: studies with the human growth hormone gene. Gene. 1986;46(2-3):277–286. doi: 10.1016/0378-1119(86)90412-9. [DOI] [PubMed] [Google Scholar]
  7. Dobb M. G., Murray R., Sikorski J. Specific labelling of thiol groups in mammalian keratin suitable for electron microscope studies. J Microsc. 1972 Dec;96(3):285–299. doi: 10.1111/j.1365-2818.1972.tb01059.x. [DOI] [PubMed] [Google Scholar]
  8. Efstratiadis A., Posakony J. W., Maniatis T., Lawn R. M., O'Connell C., Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M., Slightom J. L. The structure and evolution of the human beta-globin gene family. Cell. 1980 Oct;21(3):653–668. doi: 10.1016/0092-8674(80)90429-8. [DOI] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  10. Frenkel M. J., Powell B. C., Ward K. A., Sleigh M. J., Rogers G. E. The keratin BIIIB gene family: isolation of cDNA clones and structure of a gene and a related pseudogene. Genomics. 1989 Feb;4(2):182–191. doi: 10.1016/0888-7543(89)90298-x. [DOI] [PubMed] [Google Scholar]
  11. Gillespie J. M., Broad A. Ultra-high-sulphur proteins in the hairs of the artiodactyla. Aust J Biol Sci. 1972 Feb;25(1):138–145. [PubMed] [Google Scholar]
  12. Gillespie J. M., Marshall R. C., Moore G. P., Panaretto B. A., Robertson D. M. Changes in the proteins of wool following treatment of sheep with epidermal growth factor. J Invest Dermatol. 1982 Sep;79(3):197–200. doi: 10.1111/1523-1747.ep12500056. [DOI] [PubMed] [Google Scholar]
  13. Gillespie J. M., Reis P. J. The dietary-regulated biosynthesis of high-sulphur wool proteins. Biochem J. 1966 Mar;98(3):669–677. doi: 10.1042/bj0980669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson P. F., McKnight S. L. Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989;58:799–839. doi: 10.1146/annurev.bi.58.070189.004055. [DOI] [PubMed] [Google Scholar]
  15. Konarska M. M., Padgett R. A., Sharp P. A. Recognition of cap structure in splicing in vitro of mRNA precursors. Cell. 1984 Oct;38(3):731–736. doi: 10.1016/0092-8674(84)90268-x. [DOI] [PubMed] [Google Scholar]
  16. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kozak M. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res. 1981 Oct 24;9(20):5233–5252. doi: 10.1093/nar/9.20.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Krieg P. A., Melton D. A. In vitro RNA synthesis with SP6 RNA polymerase. Methods Enzymol. 1987;155:397–415. doi: 10.1016/0076-6879(87)55027-3. [DOI] [PubMed] [Google Scholar]
  19. Kuczek E. S., Rogers G. E. Sheep wool (glycine + tyrosine)-rich keratin genes. A family of low sequence homology. Eur J Biochem. 1987 Jul 1;166(1):79–85. doi: 10.1111/j.1432-1033.1987.tb13486.x. [DOI] [PubMed] [Google Scholar]
  20. Kuczek E., Rogers G. E. Sheep keratins: characterization of cDNA clones for the glycine + tyrosine-rich wool proteins using a synthetic probe. Eur J Biochem. 1985 Jan 2;146(1):89–93. doi: 10.1111/j.1432-1033.1985.tb08623.x. [DOI] [PubMed] [Google Scholar]
  21. McLauchlan J., Gaffney D., Whitton J. L., Clements J. B. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini. Nucleic Acids Res. 1985 Feb 25;13(4):1347–1368. doi: 10.1093/nar/13.4.1347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McNab A. R., Wood L., Theriault N., Gierman T., Vogeli G. An ultra-high sulfur keratin gene is expressed specifically during hair growth. J Invest Dermatol. 1989 Feb;92(2):263–266. doi: 10.1111/1523-1747.ep12276832. [DOI] [PubMed] [Google Scholar]
  23. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  24. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  25. Powell B. C., Rogers G. E. Cyclic hair-loss and regrowth in transgenic mice overexpressing an intermediate filament gene. EMBO J. 1990 May;9(5):1485–1493. doi: 10.1002/j.1460-2075.1990.tb08266.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Powell B. C., Sleigh M. J., Ward K. A., Rogers G. E. Mammalian keratin gene families: organisation of genes coding for the B2 high-sulphur proteins of sheep wool. Nucleic Acids Res. 1983 Aug 25;11(16):5327–5346. doi: 10.1093/nar/11.16.5327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  28. ROGERS G. E. Electron microscope studies of hair and wool. Ann N Y Acad Sci. 1959 Nov 20;83:378–399. doi: 10.1111/j.1749-6632.1960.tb40914.x. [DOI] [PubMed] [Google Scholar]
  29. ROTH S. I., HELWIG E. B. THE CYTOLOGY OF THE DERMAL PAPILLA, THE BULB, AND THE ROOT SHEATHS OF THE MOUSE HAIR. J Ultrastruct Res. 1964 Aug;11:33–51. doi: 10.1016/s0022-5320(64)80091-5. [DOI] [PubMed] [Google Scholar]
  30. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Southern E. Gel electrophoresis of restriction fragments. Methods Enzymol. 1979;68:152–176. doi: 10.1016/0076-6879(79)68011-4. [DOI] [PubMed] [Google Scholar]
  35. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  36. Winter G., Fields S. Cloning of influenza cDNA ino M13: the sequence of the RNA segment encoding the A/PR/8/34 matrix protein. Nucleic Acids Res. 1980 May 10;8(9):1965–1974. doi: 10.1093/nar/8.9.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES