Abstract
Cell and DNA replicative potentials were studied in 10 strains of skin fibroblasts from unrelated patients with the Werner syndrome (WS) and in a progeric CRL 1277 strain. The lifespans of all WS strains and of CRL 1277 cells are greatly abbreviated in vitro, due to large fractions of non-cycling cells and the basic process of progressive clonal attenuation during Phase II. In some WS strains, the population-doubling rate per day and the cloning efficiency fluctuated concurrently in random fashion during the cellular aging process, indicating alternating successions of adaptively well and poorly growing clones, probably resulting from various chromosome translocations. No detectable defect in excision repair was found in WS or CRL 1277 cells. However, the rate of increase in the molecular weight of pulse-chased DNA involving overall rates of chain elongation and replicon fusion was retarded in WS and CRL 1277 cells. Also, pulse-labelled DNA in WS fibroblasts was less enriched in the nuclear matrix and was more slowly chased out than in normal cells. These results led us to postulate a misfiring or delayed initiation due to the sticky attachment of replicating DNA to the nuclear matrix in the replisomes of WS fibroblasts. A suggested model for abnormal DNA replication is presented and discussed to explain the loss of DNA and the chromosome abnormalities in WS cells. The abnormal DNA-synthetic profiles so derived appeared to be normalized in SV40-transformed PSV811 (WS) cells as were in gamma ray-transformed wild-type WI38CT-1 cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Absher, P.M., and Absher, R.G. (1976) Clonal variation and aging of diploid fibroblasts. Exp. Cell Res., 103: 247–255.
Bell, E., Marek, L.F., Levinstone, D.S., Merril, C., Sher, S.,Young, I.T., and Eden, M. (1978) Loss of division potential in vitro: aging or differentiation. Science, 202: 1158–1163.
Cleaver, J.E. (1978) DNA repair and its coupling to DNA replication in eukaryotic cells. Biochim. Biophys. Acta, 516: 489–516.
Epstein, C.J., Martin, G.M., Schultz, A.L., and Motulsky, A.G. (1966) Werner’s syndrome: A review of its symptomatology, natural history, pathological features, genetics and relationship to the natural aging process. Medicine, 45: 177–221.
Epstein, J., Williams, J.R., and Little, J.B. (1974) Rate of DNA repair in progeric and normal human cells. Biochem. Biophys. Res. Commun., 59: 850–857.
Fujiwara, Y. (1972) Effect of cycloheximide on regulatory protein for initiating mammalian DNA replication at the nuclear membrane. Cancer Res., 32: 2089–2095.
Fujiwara, Y., Goto, K., and Kano, Y. (1982) Ultraviolet hypersensitivity of Cockayne’s syndrome fibroblasts, effects of nictoniamide adenine dinucleotide and poly (ADP-ribose) synthesis. Exp. Cell Res., 138: 207–215.
Fujiwara, Y. (1983) Roles of poly (ADP-ribose) synthesis in repair and replication in normal human, Cockayne syndrome and xeroderma pigmentosum cells after UV irradiation. In:ADP-Ribosylation, DNA Repair and Cancer, M. Miwa et al., (Eds.), Japan Sci. Soc. Press, Tokyo, pp. 209–218.
Fujiwara, Y., Higashikawa, T., and Tatsumi, M. (1977a) A retarded rate of DNA replication and normal level of DNA repair in Werner’s syndrome fibroblasts in culture. J. Cell Physiol. 92: 365–374.
Fujiwara, Y., Ichihashi, M., Kano, Y., Goto, K., and Shimize, K. (1981) A new human photosensitive subject with a defect in the recovery of DNA synthesis after UV irradiation. J. Invest. Dermatol., 77: 256–263.
Fujiwara, Y., and Tatsumi, M. (1976) Replication bypass repair of ultraviolet damage to DNA of mammalian cells: caffeine resistant and sensitive mechanisms. Mutation Res., 37: 91110.
Fujiwara, Y., Tatsumi, M., and Sasaki, M.S. (1977b) Crosslink repair in human cells and its possible defect in Fanconi’s anemia cells. J. Mol. Biol., 113: 635–649.
Fujiwara, Y., and Satoh, Y. (1981) Age-dependent change in fibroblast cultures from xeroderma pigmentosum variant. J. Invest. Dermatol. 76: 215–220.
Goldstein, S., and Moreman, E.J. (1975) Heat-labile enzymes in Werner’s syndrome fibroblasts. Nature, 255: 159.
Hart, R.W., and Setlow, R.B. (1974) Correlation between DNA excision-repair and life-span in a number of mammalian species. Proc. Nati. Acad. Sci. USA, 71: 2169–2173.
Hayflick, L. (1965) The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res., 37: 614–636.
Higashikawa, T., and Fujiwara, Y. (1978) Normal level of unscheduled DNA synthesis in Werner’s syndrome fibroblasts in culture. Exp. Cell Res., 113: 438–442.
Hoehn, H., Bryant, E.M., Au, K., Norwood, T.H., Boman, H., and Martin, G.M. (1975) Variegated translocation mosaicism in human skin fibroblast cultures. Cytogenet. Cell Genet., 15: 282–298.
Martin, G.M. (1978) Genetic syndromes in man with potential relevance to pathobiology of aging. Birth Defects Orig. Art. Ser., 14: 5–39.
Martin, G.M., Sprague, C.A., and Epstein, C.J. (1970) Replicative lifespan of cultivated human cells: effects of donor’s age, tissue and genotype. Lab. Invest., 23: 86–91.
Martin, G.M., Sprague, C.A., Norwood, T.H., and Pendergrass, W.R. (1974) Clonal selection, attenuation and differentiation in in vitro model of hyperplasia. Am. J. Pathol. 94: 137–150.
McCready, S.J., Kackson, D.A. and Cook, P.R. (1982) Attachment of intact superhelical DNA to the nuclear cage during replication and transcription. Prog. Mutat. Res. 4: 113–130.
Nakao, Y., Kishihara, M., Yoshimi, M., Inoue, Y., Tanaka, K.,Sakamoto, N., Imura, H., Ichihashi, M., and Fujiwara, Y. (1978) Werner’s syndrome: in vivo and in vitro characteristics as a model of aging. Am. J. Med., 65: 919932.
Namba, M., Nishitani, K., and Kimoto, T. (1980) Characteristics of WI-38 cells (WI38CT-1) transformed by treatment with Co-60 gamma rays. Gann, 71: 300–307.
Norwood, T.H., Hoehn, H., Salk, D., Martin, G.M. (1979) Cellular aging in Werner’s syndrome: a unique phenotype? J. Invest. Dermatol., 73: 92–96.
Norwood, T.H., Pendergrass, W.R., Sprague, C.A., and Martin, G.M. (1974) Dominance of the senescent phenotype in the heterokaryon between replicative and post-replicative human fibroblast-like cells. Proc. Natl. Acad. Sci. USA, 71: 2231–2235.
Ohashi, M., Aizawa, Ooka, H., Ohsawa, T., Kaji, K., Kondo, H. and T. Utakoji (1980) A new human diploid cell strain, TIG-1, for the research on cellular aging. Exp. Gerontol., 15: 121133.
Pardoll, D.W., Vogelstein, B., and Coffey, D.S. (1980) A fixed site of DNA replication in eukaryotic cells., Cell 19: 527536.
Regan, J.D., and Setlow, R.B. (1974) DNA repair in human progeroid cells. Biochem. Biophys. Res. Commun., 59: 858864.
Salk, D. (1982) Werner’s syndrome: a review of recent research with analysis of connective tissue metabolism, growth control of cultured cells and chromosomal aberrations. Human Genet., 62: 1–15.
Salk, D., Au, K., Hoehn, H., and Martin, G.M. (1981a) Cytogenetics of Werner’s syndrome cultured skin fibroblasts: Variegated translocation mosaicism. Cytogenet. Cell Genet., 30: 92–107.
Salk, D., Au, K., Hoehn, H., Stenchever, M.R., and Martin, G.M. (1981b) Evidence of clonal attenuation, clonal succession and clonal expansion in mass cultures of aging Werner’s syndrome skin fibroblasts. Cytogenet. Cell Genet., 30: 108–117.
Schneider, E.L., and Mitsui, Y. (1976) The relationship between in vitro cellular aging and in vivo human aging. Proc. Natl. Acad. Sci. USA, 73: 3584–3588.
Smith, J.R., Pereira-Smith, O.M., and Schneider, E.L. (1978)Colony size distributions as a measure of in vivo and in vitro aging. Proc. Natl. Acad. Sci. USA, 75: 1353–1356.
Smith, J.R., and Whitney, R.G. (1980) Interclonal ariation in proliferative potential of human diploid fibroblasts:stochastic mechanism for cellular aging. Nature, 207: 82–84.
Takeuchi, F., Hanaoka, F., Goto, M., Akaoka, I., Hori, T.,Yamada, M., and Miyamoto, T. (1982) Altered frequency of initiation sites of DNA replication in Werner’s syndrome cells. Human Genet., 60: 365–368.
Tanaka, K., Nakazawa, T., Okada, Y., and Kumahara, Y. (1979) Increase in Werner’s syndrome cells by hybridization with normal human diploid and HeLa cells. Exp. Cell Res., 123: 261–267.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1985 Plenum Press, New York
About this chapter
Cite this chapter
Fujiwara, Y., Kano, Y., Ichihashi, M., Nakao, Y., Matsumura, T. (1985). Abnormal Fibroblast Aging and DNA Replication in the Werner Syndrome. In: Salk, D., Fujiwara, Y., Martin, G.M. (eds) Werner’s Syndrome and Human Aging. Advances in Experimental Medicine and Biology, vol 190. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-7853-2_23
Download citation
DOI: https://doi.org/10.1007/978-1-4684-7853-2_23
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-7855-6
Online ISBN: 978-1-4684-7853-2
eBook Packages: Springer Book Archive