• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. May 1992; 11(5): 1921–1929.
PMCID: PMC556651

Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity.

Abstract

Loss of telomeric DNA during cell proliferation may play a role in ageing and cancer. Since telomeres permit complete replication of eukaryotic chromosomes and protect their ends from recombination, we have measured telomere length, telomerase activity and chromosome rearrangements in human cells before and after transformation with SV40 or Ad5. In all mortal populations, telomeres shortened by approximately 65 bp/generation during the lifespan of the cultures. When transformed cells reached crisis, the length of the telomeric TTAGGG repeats was only approximately 1.5 kbp and many dicentric chromosomes were observed. In immortal cells, telomere length and frequency of dicentric chromosomes stabilized after crisis. Telomerase activity was not detectable in control or extended lifespan populations but was present in immortal populations. These results suggest that chromosomes with short (TTAGGG)n tracts are recombinogenic, critically shortened telomeres may be incompatible with cell proliferation and stabilization of telomere length by telomerase may be required for immortalization.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allshire RC, Gosden JR, Cross SH, Cranston G, Rout D, Sugawara N, Szostak JW, Fantes PA, Hastie ND. Telomeric repeat from T. thermophila cross hybridizes with human telomeres. Nature. 1988 Apr 14;332(6165):656–659. [PubMed]
  • Allshire RC, Dempster M, Hastie ND. Human telomeres contain at least three types of G-rich repeat distributed non-randomly. Nucleic Acids Res. 1989 Jun 26;17(12):4611–4627. [PMC free article] [PubMed]
  • Bauer GA, Heller HM, Burgers PM. DNA polymerase III from Saccharomyces cerevisiae. I. Purification and characterization. J Biol Chem. 1988 Jan 15;263(2):917–924. [PubMed]
  • Benn PA. Specific chromosome aberrations in senescent fibroblast cell lines derived from human embryos. Am J Hum Genet. 1976 Sep;28(5):465–473. [PMC free article] [PubMed]
  • Blackburn EH. Structure and function of telomeres. Nature. 1991 Apr 18;350(6319):569–573. [PubMed]
  • Blackburn EH. The molecular structure of centromeres and telomeres. Annu Rev Biochem. 1984;53:163–194. [PubMed]
  • Brown WR. Molecular cloning of human telomeres in yeast. Nature. 1989 Apr 27;338(6218):774–776. [PubMed]
  • Chang SE. In vitro transformation of human epithelial cells. Biochim Biophys Acta. 1986;823(3):161–194. [PubMed]
  • Cooke HJ, Brown WR, Rappold GA. Hypervariable telomeric sequences from the human sex chromosomes are pseudoautosomal. Nature. 1985 Oct 24;317(6039):687–692. [PubMed]
  • Cross SH, Allshire RC, McKay SJ, McGill NI, Cooke HJ. Cloning of human telomeres by complementation in yeast. Nature. 1989 Apr 27;338(6218):771–774. [PubMed]
  • de Lange T, Shiue L, Myers RM, Cox DR, Naylor SL, Killery AM, Varmus HE. Structure and variability of human chromosome ends. Mol Cell Biol. 1990 Feb;10(2):518–527. [PMC free article] [PubMed]
  • DiPaolo JA. Relative difficulties in transforming human and animal cells in vitro. J Natl Cancer Inst. 1983 Jan;70(1):3–8. [PubMed]
  • Fitzgerald PH, Morris CM. Telomeric association of chromosomes in B-cell lymphoid leukemia. Hum Genet. 1984;67(4):385–390. [PubMed]
  • Goldstein S. Replicative senescence: the human fibroblast comes of age. Science. 1990 Sep 7;249(4973):1129–1133. [PubMed]
  • Graham FL. Growth of 293 cells in suspension culture. J Gen Virol. 1987 Mar;68(Pt 3):937–940. [PubMed]
  • Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. [PubMed]
  • Graham FL, Smiley J, Russell WC, Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. [PubMed]
  • Greider CW, Blackburn EH. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell. 1985 Dec;43(2 Pt 1):405–413. [PubMed]
  • Greider CW, Blackburn EH. The telomere terminal transferase of Tetrahymena is a ribonucleoprotein enzyme with two kinds of primer specificity. Cell. 1987 Dec 24;51(6):887–898. [PubMed]
  • Greider CW, Blackburn EH. A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature. 1989 Jan 26;337(6205):331–337. [PubMed]
  • Harley CB. Telomere loss: mitotic clock or genetic time bomb? Mutat Res. 1991 Mar-Nov;256(2-6):271–282. [PubMed]
  • Harley CB, Futcher AB, Greider CW. Telomeres shorten during ageing of human fibroblasts. Nature. 1990 May 31;345(6274):458–460. [PubMed]
  • Hastie ND, Dempster M, Dunlop MG, Thompson AM, Green DK, Allshire RC. Telomere reduction in human colorectal carcinoma and with ageing. Nature. 1990 Aug 30;346(6287):866–868. [PubMed]
  • Lundblad V, Szostak JW. A mutant with a defect in telomere elongation leads to senescence in yeast. Cell. 1989 May 19;57(4):633–643. [PubMed]
  • Meisner LF, Wu SQ, Christian BJ, Reznikoff CA. Cytogenetic instability with balanced chromosome changes in an SV40 transformed human uroepithelial cell line. Cancer Res. 1988 Jun 1;48(11):3215–3220. [PubMed]
  • Meyne J, Ratliff RL, Moyzis RK. Conservation of the human telomere sequence (TTAGGG)n among vertebrates. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7049–7053. [PMC free article] [PubMed]
  • MOORHEAD PS, SAKSELA E. NON-RANDOM CHROMOSOMAL ABERRATIONS IN SV 40-TRANSFORMED HUMAN CELLS. J Cell Physiol. 1963 Aug;62:57–83. [PubMed]
  • Moorhead PS, Saksela E. The sequence of chromosome aberrations during SV 40 transformation of a human diploid cell strain. Hereditas. 1965;52(3):271–284. [PubMed]
  • Morgan AR, Lee JS, Pulleyblank DE, Murray NL, Evans DH. Review: ethidium fluorescence assays. Part 1. Physicochemical studies. Nucleic Acids Res. 1979 Oct 10;7(3):547–569. [PMC free article] [PubMed]
  • Morin GB. The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell. 1989 Nov 3;59(3):521–529. [PubMed]
  • Moyzis RK, Buckingham JM, Cram LS, Dani M, Deaven LL, Jones MD, Meyne J, Ratliff RL, Wu JR. A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6622–6626. [PMC free article] [PubMed]
  • Olovnikov AM. A theory of marginotomy. The incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon. J Theor Biol. 1973 Sep 14;41(1):181–190. [PubMed]
  • Pathak S, Wang Z, Dhaliwal MK, Sacks PC. Telomeric association: another characteristic of cancer chromosomes? Cytogenet Cell Genet. 1988;47(4):227–229. [PubMed]
  • Radna RL, Caton Y, Jha KK, Kaplan P, Li G, Traganos F, Ozer HL. Growth of immortal simian virus 40 tsA-transformed human fibroblasts is temperature dependent. Mol Cell Biol. 1989 Jul;9(7):3093–3096. [PMC free article] [PubMed]
  • Sack GH., Jr Human cell transformation by simian virus 40--a review. In Vitro. 1981 Jan;17(1):1–19. [PubMed]
  • SAKSELA E, MOORHEAD PS. ANEUPLOIDY IN THE DEGENERATIVE PHASE OF SERIAL CULTIVATION OF HUMAN CELL STRAINS. Proc Natl Acad Sci U S A. 1963 Aug;50:390–395. [PMC free article] [PubMed]
  • Shay JW, Wright WE. Quantitation of the frequency of immortalization of normal human diploid fibroblasts by SV40 large T-antigen. Exp Cell Res. 1989 Sep;184(1):109–118. [PubMed]
  • Sherwood SW, Rush D, Ellsworth JL, Schimke RT. Defining cellular senescence in IMR-90 cells: a flow cytometric analysis. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9086–9090. [PMC free article] [PubMed]
  • Shippen-Lentz D, Blackburn EH. Telomere terminal transferase activity from Euplotes crassus adds large numbers of TTTTGGGG repeats onto telomeric primers. Mol Cell Biol. 1989 Jun;9(6):2761–2764. [PMC free article] [PubMed]
  • Southern PJ, Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed]
  • Spanos A, Sedgwick SG, Yarranton GT, Hübscher U, Banks GR. Detection of the catalytic activities of DNA polymerases and their associated exonucleases following SDS-polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Apr 24;9(8):1825–1839. [PMC free article] [PubMed]
  • Stampfer M, Hallowes RC, Hackett AJ. Growth of normal human mammary cells in culture. In Vitro. 1980 May;16(5):415–425. [PubMed]
  • Stein GH. SV40-transformed human fibroblasts: evidence for cellular aging in pre-crisis cells. J Cell Physiol. 1985 Oct;125(1):36–44. [PubMed]
  • Stewart N, Bacchetti S. Expression of SV40 large T antigen, but not small t antigen, is required for the induction of chromosomal aberrations in transformed human cells. Virology. 1991 Jan;180(1):49–57. [PubMed]
  • Walen KH. Chromosome instability in cell lineages of amniocyte clones morphologically transformed by Simian virus 40. Cancer Genet Cytogenet. 1987 Mar;25(1):149–159. [PubMed]
  • Yudkin J. Sugar and disease. Nature. 1972 Sep 22;239(5369):197–199. [PubMed]
  • WOLMAN SR, HIRSCHHORN K, TODARO GJ. EARLY CHROMOSOMAL CHANGES IN SV40-INFECTED HUMAN FIBROBLAST CULTURES. Cytogenetics. 1964;3:45–61. [PubMed]
  • Wright WE, Pereira-Smith OM, Shay JW. Reversible cellular senescence: implications for immortalization of normal human diploid fibroblasts. Mol Cell Biol. 1989 Jul;9(7):3088–3092. [PMC free article] [PubMed]
  • Yu GL, Bradley JD, Attardi LD, Blackburn EH. In vivo alteration of telomere sequences and senescence caused by mutated Tetrahymena telomerase RNAs. Nature. 1990 Mar 8;344(6262):126–132. [PubMed]
  • Zahler AM, Prescott DM. Telomere terminal transferase activity in the hypotrichous ciliate Oxytricha nova and a model for replication of the ends of linear DNA molecules. Nucleic Acids Res. 1988 Jul 25;16(14B):6953–6972. [PMC free article] [PubMed]
  • Zakian VA. Structure and function of telomeres. Annu Rev Genet. 1989;23:579–604. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...