Histone H1 in G1 Arrested, Senescent, and Werner Syndrome Fibroblasts

  • Youji Mitsui
  • Hiroshi Sakagami
  • Masa-atu Yamada
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 190)


Histone H1 content and synthesis were examined in normal, Werner-syndrome, and transformed fibroblasts. Analysis of 3H-lysine incorporation indicated that senescent cells, but not G1-arrested young cells, had a lower ratio of molar synthesis of H1 histone to nucleosome histones than did growing young cells or gamma-ray-transformed cells. Furthermore, a biochemical study of histone H1 content plotted as a function of DNA synthesis activity and an immunocytological study using antiserum against histone Hi revealed that senescent cells had a lower histone H1 content than did young cultures at all stages of cell proliferation. Werner syndrome skin fibroblasts at early passage, however, had amounts of histone Hi comparable to those of age-matched normal control fibroblasts. We conclude that a decline, with increasing passage number, in content and synthesis of H1 histone relative to nucleosomal histones (Mitsui et al., 1980) was not simply due to passage-related accumulation of G1-arrested cells, but actually reflected age specific changes of cultured human fibroblasts. The depletion of histone H1 in the chromatin of senescent cells is a possible cause of DNA strand breakage or relaxation of gene repression.


Senescent Cell Young Cell WERNER Syndrome Human Diploid Fibroblast Population Doubling Level 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aizawa, S. and Mitsui, Y. (1979), A new cell surface marker of aging in human diploid fibroblasts, J. Cell. Physiol 100:383.Google Scholar
  2. Aizawa, S., Mitsui, Y., Kurimoto, F. and Matuoka, K. (1980), Cell surface changes accompanying aging in human diploid fibroblasts. III. Division age and senescence revealed by concanavaline A-mediated red blood cell adsorption, Expl Cell Res 125: 297.Google Scholar
  3. Appels, R., Bolund, L. and Ringertz, N.R. (1974), Biochemical analysis of reactiviated chick erythrocyte nuclei isolated from chick/HeLa heterokaryons, J. Mol. Biol 87:339.Google Scholar
  4. Beaupain, R., Icard, C., and Maciera-Coelho, A. (1980), Changes inGoogle Scholar
  5. DNA alkalin-sensitive sites during senescence and establishment of fibroblasts in vitro, Biochim Biophys Acta 606:251.Google Scholar
  6. Berlowitz, E.M. and Doty, P. (1975), Chemical and physical properties of fractionated chromatin, Proc. Natl. Acad Sci. USA, 72:328.Google Scholar
  7. Berger, N.A., Petzold, S.J. and Berger, S.J. (1979), Association of poly (ADP-rib) synthesis with cessation of DNA synthesis and DNA fragmentation. Biochim Biophys Acta 564: 90.Google Scholar
  8. Bowman, P.D., Meek, R.L. and Daniel, C.W. (1975), Aging of human fibroblasts in vitro., Exp. Cell Res. 93:184.Google Scholar
  9. Chiu, N., Baserga, R. and Furth, J.J., (1977), Composition and template activity of chromatin fractionated by isoelectric focusing, Biochemistry 16: 4796.Google Scholar
  10. Christofalo, V.J. and Sharf, B.B. (1973), Cellular senescence and DNA synthesis, Exp Cell Res. 76: 419.Google Scholar
  11. D’Anna, J., Gurley, L. and Tobey, R., (1982), Synthesis and modulations in the chromatin contents of histones H1 during G1 and S phases in Chinese hamster cells, Biochem 21: 3991.Google Scholar
  12. Elgins, S.C.R. and Hood, L.E. (1973), Chromosomal Proteins of Drosophila, Biochem 12, 4984.Google Scholar
  13. Elgins, S.C.R. and Weintraub, H. (1975), Chromosomal proteins and chromatin structure, Annu Rev. Biochem 44: 725.Google Scholar
  14. Greenaway, P.J. and Murray, K. (1971), Heterogeneity and polymorphism in chicken erythrocyte histone fraction V. Nature 229: 233.Google Scholar
  15. Grimes, S.R. Jr., Chae, C-B and Irvin, J.L. (1975), Effects of age and hypophysectomy upon relative proportions of various histones in rat testis, Biochem. Biophys. Res. Commun 64:911.Google Scholar
  16. Gorovsky, M.A. and Keevert, J.B. (1975), Absence of histone F1 in a mitotically dividing, genetically inactive nucleus, Proc. Natl. Acad Sci. USA, 72:2672.Google Scholar
  17. Hart, R.W. and Setlow, R.B. (1976), DNA repair in late-passage human cells, Mech. Ageing Dev., 5:67.Google Scholar
  18. Herve, B., Jacquemin, E. and Courtois, Y. (1979), Histones biosynthesis and turnover in epithelial lens cells cultured in vitro, Cell. Biol. Intern. Rep 3:271.Google Scholar
  19. Houck, J.C., Sharma, U.K. and Hayflick, L. (1972), Functional failures of cultured human diploid fibroblasts after continued population doublings, Proc Soc. Exp. Biol. Med 137:331.Google Scholar
  20. Kraus, M.O. and Stein, G.S. (1974), Modifications in the chromosomal proteins of SV-40 transformed WI-38 human diploid fibroblasts, Biochem. Biophys Res. Commun 59: 796.Google Scholar
  21. Linder, S., Zuckerman, S. and Ringertz, N. (1982), Distribution of histone H5 in chicken erythrocyte-mammalian cell heterokaryons, Exp. Cell Res., 140: 464.Google Scholar
  22. Lohr, D., Tatchell, K. and Van Holde, K.E. (1977), On the occurrence of nucleosome phasing in chromatin, Cell 12: 829.Google Scholar
  23. Macieira-Coelho, A. (1974), Are non-diving cells present in ageing cell cultures? Nature 248: 421.Google Scholar
  24. Martin, G.M. (1978), Genetic syndromes in man with potential relevance to the pathology of aging, Birth Defects, Orig. Article Series 14:5.Google Scholar
  25. Mitsui, Y. and Schneider, E.L. (1976), Increased nuclear sizes in senescent diploid fibroblast cultures, Exp Cell Res 100: 147.Google Scholar
  26. Mitsui, Y. and Schneider, E.L.(1976), Characterization of fractionated human diploid fibroblast cell populations, Exp. Cell Res 103:23.Google Scholar
  27. Mitsui, Y. and Schneider, E.L.(1976), Relationship between replication and volume in senescent human diploid fibroblasts, Mech. Ageing Dev 5:45.Google Scholar
  28. Mitsui, Y., Aizawa, S. and Matuoka, K. (1979), The relation of cell nuclei and surface membranes to the capacity of cell proliferation in human diploid fibroblasts. In Recent Advances in Gerontology Orimo, H., Shimada, K., Iriki, M. andGoogle Scholar
  29. Maeda, D., editors, Excerpta Medica, Amsterdam, 108–110.Google Scholar
  30. Mitsui, Y., Sakagami, H, Murata, S. and Yamamada, M. (1980), Age related decline in H1 histone fraction in human diploid fibroblast cultures, Exp Cell Res., 126: 289.Google Scholar
  31. Mitsui, Y. Matuoka, K., Aizawa, S. and Noda, K. (1980), New approaches to the characterization of aging human diploid fibroblasts at individual cell level, Adv. Exp. Med. Biol 129: 5.Google Scholar
  32. Namba, M., Nishitani, K. and Kimoto, T., (1978), Carcinogenesis in tissue culture 29: Neoplastic transformation of a normal human diploid cell strain, WI-38, with Co-60 gamma rays, Japan J. Exp. Med 48:303.Google Scholar
  33. O’Farrell, P.H. (1975), High resolution two-dimensional electrophoresis of proteins, J. Biol. Chem 250:4007.Google Scholar
  34. Ohashi, M., Aizawa, S., Ooka, H., Ohsawa, T., Kaji, K., Kondo, H, Kobayashi, T., Noumura, T., Matsuo, M., Mitsui, Y., Murata, S., Yamamoto, K., Ito, H., Shimada,H. and Utakoji, T. (1980), A new human diploid cell strain, TIG-1, for the research on cellular aging, Exp. Gerontol 15:121.Google Scholar
  35. Ono, T., Okada, S. and Sugahara, T. (1976), Comparative studies of DNA size in various tissues of mice during the aging process, Exp. Gerontol 11:127.Google Scholar
  36. Ono, T. and Cutler, R.G. (1978), Age-dependent relaxation of repression: Increase endogenous murine leukemia virus-related and globin-related RNA in brain and liver of mice, Proc. Natl. Acad Sci. USA, 75:4431.Google Scholar
  37. Panyim, S. and Chalkley, R. (1969), High resolution acrylamide gel electrophoresis of histones, Arch. Biochem. Biophys 130:337.Google Scholar
  38. Pehrson, J. and Cole, D.R. (1980), Histone H1 accumulates in growth-inhibited cultured cells, Nature 285: 43.PubMedCrossRefGoogle Scholar
  39. Peng, M.T. and Lee, L.R. (1979), Regional differences of neuron loss of rat brain in old age, Gerontology 25: 205.Google Scholar
  40. Pochran, S.F., Omeara, A.R and Kurtz, M.J. (1978), Control of transcription in ageing W1–38 cells stimulated to divide, Exp. Cell Res., 116:63.Google Scholar
  41. Price, G.B., Modak, S.D. and Makinodan, T. (1971), Age-associated changes in the DNA of mouse tissue, Science, 171: 917.PubMedCrossRefGoogle Scholar
  42. Sakagami, H., Mitsui, Y., Murata, S. and Yamada, M. (1982), Effect of growth stage on histone H1 metabolism in human diploid fibroblasts, J. Cell Physiol 110: 213.PubMedCrossRefGoogle Scholar
  43. Salk, D., Au, K., Hoehn, H. and Martin, G.M. (1981), Cytogenetics of Werner syndrome cultured skin fibroblasts: Variegated translocation mosaicism, Cytogenet. Cell Genet., 30:92Google Scholar
  44. Schneider, E.L., Mitsui, Y., Tice, R., Shorr, S.S. and Braunschweiger, K. (1975), Alteration in cellular RNAs during the in vitro lifespan of cultured human diploid fibroblasts, Mech. Ageing Dev 4:449.Google Scholar
  45. Seale, R.L. and Alonson, A.I. (1973), Chromatin-associated proteins of the developing sea-urchin embryo. 11 Acid-soluble proteins, J. Mol. Biol 75: 647.Google Scholar
  46. Shirley, M.A and Anderson, K.M. (1977), Electron-microscopic visualization of transcriptionally active and less active chromatin fractions from the rat ventral prostate and their content of histones, Can. J. Biochem 55:9.Google Scholar
  47. Takeuchi, F., Hanaoka, F., Goto, M., Yamada, M. and Miyamoto, T. (1982), Prolongation of S phase and whole cell cycle in Werner syndrome fibroblasts, Exp. Geront 17:473.Google Scholar
  48. Tarnowka, M.A., Baglioni, C. and Basilico, C. (1978), Synthesis of H1 histones by BHK cells in G1, Cell 15: 163.Google Scholar
  49. Worcel, Z. and Benyajuti, C. (1977), Higher order coiling of DNA chromatin, Cell 12: 83.Google Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Youji Mitsui
    • 1
  • Hiroshi Sakagami
    • 1
  • Masa-atu Yamada
    • 1
  1. 1.Division of Cell Science and Technology, Agency of Industrial Science and TechnologyFermentation Research InstituteYatabe-machi, Ibaraki, 305Japan

Personalised recommendations