Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1999 Jun 15;18(12):3509–3519. doi: 10.1093/emboj/18.12.3509

Progressive cis-inhibition of telomerase upon telomere elongation.

S Marcand 1, V Brevet 1, E Gilson 1
PMCID: PMC1171430  PMID: 10369690

Abstract

In yeast, the constant length of telomeric DNA results from a negative regulation of telomerase by the telomere itself. Here we follow the return to equilibrium of an abnormally shortened telomere. We observe that telomere elongation is restricted to a few base pairs per generation and that its rate decreases progressively with increasing telomere length. In contrast, in the absence of telomerase or in the presence of an over-elongated telomere, the degradation rate linked to the succession of generations appears to be constant, i.e. independent of telomere length. Together, these results indicate that telomerase is gradually inhibited at its site of action by the elongating telomere. The implications of this finding for the dynamics of telomere length regulation are discussed in this study.

Full Text

The Full Text of this article is available as a PDF (381.0 KB).

Selected References

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

  1. Aparicio O. M., Billington B. L., Gottschling D. E. Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae. Cell. 1991 Sep 20;66(6):1279–1287. doi: 10.1016/0092-8674(91)90049-5. [DOI] [PubMed] [Google Scholar]
  2. Barnett M. A., Buckle V. J., Evans E. P., Porter A. C., Rout D., Smith A. G., Brown W. R. Telomere directed fragmentation of mammalian chromosomes. Nucleic Acids Res. 1993 Jan 11;21(1):27–36. doi: 10.1093/nar/21.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bilaud T., Brun C., Ancelin K., Koering C. E., Laroche T., Gilson E. Telomeric localization of TRF2, a novel human telobox protein. Nat Genet. 1997 Oct;17(2):236–239. doi: 10.1038/ng1097-236. [DOI] [PubMed] [Google Scholar]
  4. Bilaud T., Koering C. E., Binet-Brasselet E., Ancelin K., Pollice A., Gasser S. M., Gilson E. The telobox, a Myb-related telomeric DNA binding motif found in proteins from yeast, plants and human. Nucleic Acids Res. 1996 Apr 1;24(7):1294–1303. doi: 10.1093/nar/24.7.1294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blue C., Marcand S., Gilson E. Proteins that bind to double-stranded regions of telomeric DNA. Trends Cell Biol. 1997 Aug;7(8):317–324. doi: 10.1016/S0962-8924(97)01092-1. [DOI] [PubMed] [Google Scholar]
  6. Boulton S. J., Jackson S. P. Identification of a Saccharomyces cerevisiae Ku80 homologue: roles in DNA double strand break rejoining and in telomeric maintenance. Nucleic Acids Res. 1996 Dec 1;24(23):4639–4648. doi: 10.1093/nar/24.23.4639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Broccoli D., Smogorzewska A., Chong L., de Lange T. Human telomeres contain two distinct Myb-related proteins, TRF1 and TRF2. Nat Genet. 1997 Oct;17(2):231–235. doi: 10.1038/ng1097-231. [DOI] [PubMed] [Google Scholar]
  8. Chong L., van Steensel B., Broccoli D., Erdjument-Bromage H., Hanish J., Tempst P., de Lange T. A human telomeric protein. Science. 1995 Dec 8;270(5242):1663–1667. doi: 10.1126/science.270.5242.1663. [DOI] [PubMed] [Google Scholar]
  9. Conrad M. N., Wright J. H., Wolf A. J., Zakian V. A. RAP1 protein interacts with yeast telomeres in vivo: overproduction alters telomere structure and decreases chromosome stability. Cell. 1990 Nov 16;63(4):739–750. doi: 10.1016/0092-8674(90)90140-a. [DOI] [PubMed] [Google Scholar]
  10. Cooper J. P., Nimmo E. R., Allshire R. C., Cech T. R. Regulation of telomere length and function by a Myb-domain protein in fission yeast. Nature. 1997 Feb 20;385(6618):744–747. doi: 10.1038/385744a0. [DOI] [PubMed] [Google Scholar]
  11. Dunn B., Szauter P., Pardue M. L., Szostak J. W. Transfer of yeast telomeres to linear plasmids by recombination. Cell. 1984 Nov;39(1):191–201. doi: 10.1016/0092-8674(84)90205-8. [DOI] [PubMed] [Google Scholar]
  12. Froelich-Ammon S. J., Dickinson B. A., Bevilacqua J. M., Schultz S. C., Cech T. R. Modulation of telomerase activity by telomere DNA-binding proteins in Oxytricha. Genes Dev. 1998 May 15;12(10):1504–1514. doi: 10.1101/gad.12.10.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gilson E., Laroche T., Gasser S. M. Telomeres and the functional architecture of the nucleus. Trends Cell Biol. 1993 Apr;3(4):128–134. doi: 10.1016/0962-8924(93)90175-z. [DOI] [PubMed] [Google Scholar]
  14. Gilson E., Roberge M., Giraldo R., Rhodes D., Gasser S. M. Distortion of the DNA double helix by RAP1 at silencers and multiple telomeric binding sites. J Mol Biol. 1993 May 20;231(2):293–310. doi: 10.1006/jmbi.1993.1283. [DOI] [PubMed] [Google Scholar]
  15. Gottschling D. E., Aparicio O. M., Billington B. L., Zakian V. A. Position effect at S. cerevisiae telomeres: reversible repression of Pol II transcription. Cell. 1990 Nov 16;63(4):751–762. doi: 10.1016/0092-8674(90)90141-z. [DOI] [PubMed] [Google Scholar]
  16. Gravel S., Larrivée M., Labrecque P., Wellinger R. J. Yeast Ku as a regulator of chromosomal DNA end structure. Science. 1998 May 1;280(5364):741–744. doi: 10.1126/science.280.5364.741. [DOI] [PubMed] [Google Scholar]
  17. Greider C. W. Telomere length regulation. Annu Rev Biochem. 1996;65:337–365. doi: 10.1146/annurev.bi.65.070196.002005. [DOI] [PubMed] [Google Scholar]
  18. Holmes S. G., Broach J. R. Silencers are required for inheritance of the repressed state in yeast. Genes Dev. 1996 Apr 15;10(8):1021–1032. doi: 10.1101/gad.10.8.1021. [DOI] [PubMed] [Google Scholar]
  19. Kennedy B. K., Austriaco N. R., Jr, Zhang J., Guarente L. Mutation in the silencing gene SIR4 can delay aging in S. cerevisiae. Cell. 1995 Feb 10;80(3):485–496. doi: 10.1016/0092-8674(95)90499-9. [DOI] [PubMed] [Google Scholar]
  20. Kimmerly W. J., Rine J. Replication and segregation of plasmids containing cis-acting regulatory sites of silent mating-type genes in Saccharomyces cerevisiae are controlled by the SIR genes. Mol Cell Biol. 1987 Dec;7(12):4225–4237. doi: 10.1128/mcb.7.12.4225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klein F., Laroche T., Cardenas M. E., Hofmann J. F., Schweizer D., Gasser S. M. Localization of RAP1 and topoisomerase II in nuclei and meiotic chromosomes of yeast. J Cell Biol. 1992 Jun;117(5):935–948. doi: 10.1083/jcb.117.5.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Krauskopf A., Blackburn E. H. Control of telomere growth by interactions of RAP1 with the most distal telomeric repeats. Nature. 1996 Sep 26;383(6598):354–357. doi: 10.1038/383354a0. [DOI] [PubMed] [Google Scholar]
  23. Krauskopf A., Blackburn E. H. Rap1 protein regulates telomere turnover in yeast. Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12486–12491. doi: 10.1073/pnas.95.21.12486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kyrion G., Boakye K. A., Lustig A. J. C-terminal truncation of RAP1 results in the deregulation of telomere size, stability, and function in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Nov;12(11):5159–5173. doi: 10.1128/mcb.12.11.5159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LaBranche H., Dupuis S., Ben-David Y., Bani M. R., Wellinger R. J., Chabot B. Telomere elongation by hnRNP A1 and a derivative that interacts with telomeric repeats and telomerase. Nat Genet. 1998 Jun;19(2):199–202. doi: 10.1038/575. [DOI] [PubMed] [Google Scholar]
  26. Lansdorp P. M., Verwoerd N. P., van de Rijke F. M., Dragowska V., Little M. T., Dirks R. W., Raap A. K., Tanke H. J. Heterogeneity in telomere length of human chromosomes. Hum Mol Genet. 1996 May;5(5):685–691. doi: 10.1093/hmg/5.5.685. [DOI] [PubMed] [Google Scholar]
  27. Larson D. D., Spangler E. A., Blackburn E. H. Dynamics of telomere length variation in Tetrahymena thermophila. Cell. 1987 Jul 31;50(3):477–483. doi: 10.1016/0092-8674(87)90501-0. [DOI] [PubMed] [Google Scholar]
  28. Lendvay T. S., Morris D. K., Sah J., Balasubramanian B., Lundblad V. Senescence mutants of Saccharomyces cerevisiae with a defect in telomere replication identify three additional EST genes. Genetics. 1996 Dec;144(4):1399–1412. doi: 10.1093/genetics/144.4.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Li B., Lustig A. J. A novel mechanism for telomere size control in Saccharomyces cerevisiae. Genes Dev. 1996 Jun 1;10(11):1310–1326. doi: 10.1101/gad.10.11.1310. [DOI] [PubMed] [Google Scholar]
  30. Lingner J., Cooper J. P., Cech T. R. Telomerase and DNA end replication: no longer a lagging strand problem? Science. 1995 Sep 15;269(5230):1533–1534. doi: 10.1126/science.7545310. [DOI] [PubMed] [Google Scholar]
  31. Lingner J., Hughes T. R., Shevchenko A., Mann M., Lundblad V., Cech T. R. Reverse transcriptase motifs in the catalytic subunit of telomerase. Science. 1997 Apr 25;276(5312):561–567. doi: 10.1126/science.276.5312.561. [DOI] [PubMed] [Google Scholar]
  32. Liu C., Mao X., Lustig A. J. Mutational analysis defines a C-terminal tail domain of RAP1 essential for Telomeric silencing in Saccharomyces cerevisiae. Genetics. 1994 Dec;138(4):1025–1040. doi: 10.1093/genetics/138.4.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lundblad V., Szostak J. W. A mutant with a defect in telomere elongation leads to senescence in yeast. Cell. 1989 May 19;57(4):633–643. doi: 10.1016/0092-8674(89)90132-3. [DOI] [PubMed] [Google Scholar]
  34. Lustig A. J. Hoogsteen G-G base pairing is dispensable for telomere healing in yeast. Nucleic Acids Res. 1992 Jun 25;20(12):3021–3028. doi: 10.1093/nar/20.12.3021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lustig A. J., Kurtz S., Shore D. Involvement of the silencer and UAS binding protein RAP1 in regulation of telomere length. Science. 1990 Oct 26;250(4980):549–553. doi: 10.1126/science.2237406. [DOI] [PubMed] [Google Scholar]
  36. Makarov V. L., Hirose Y., Langmore J. P. Long G tails at both ends of human chromosomes suggest a C strand degradation mechanism for telomere shortening. Cell. 1997 Mar 7;88(5):657–666. doi: 10.1016/s0092-8674(00)81908-x. [DOI] [PubMed] [Google Scholar]
  37. Marcand S., Buck S. W., Moretti P., Gilson E., Shore D. Silencing of genes at nontelomeric sites in yeast is controlled by sequestration of silencing factors at telomeres by Rap 1 protein. Genes Dev. 1996 Jun 1;10(11):1297–1309. doi: 10.1101/gad.10.11.1297. [DOI] [PubMed] [Google Scholar]
  38. Marcand S., Gasser S. M., Gilson E. Chromatin: a sticky silence. Curr Biol. 1996 Oct 1;6(10):1222–1225. doi: 10.1016/s0960-9822(96)00701-4. [DOI] [PubMed] [Google Scholar]
  39. Marcand S., Gilson E., Shore D. A protein-counting mechanism for telomere length regulation in yeast. Science. 1997 Feb 14;275(5302):986–990. doi: 10.1126/science.275.5302.986. [DOI] [PubMed] [Google Scholar]
  40. McEachern M. J., Blackburn E. H. Cap-prevented recombination between terminal telomeric repeat arrays (telomere CPR) maintains telomeres in Kluyveromyces lactis lacking telomerase. Genes Dev. 1996 Jul 15;10(14):1822–1834. doi: 10.1101/gad.10.14.1822. [DOI] [PubMed] [Google Scholar]
  41. McEachern M. J., Blackburn E. H. Runaway telomere elongation caused by telomerase RNA gene mutations. Nature. 1995 Aug 3;376(6539):403–409. doi: 10.1038/376403a0. [DOI] [PubMed] [Google Scholar]
  42. Murray A. W., Claus T. E., Szostak J. W. Characterization of two telomeric DNA processing reactions in Saccharomyces cerevisiae. Mol Cell Biol. 1988 Nov;8(11):4642–4650. doi: 10.1128/mcb.8.11.4642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Nakamura T. M., Cech T. R. Reversing time: origin of telomerase. Cell. 1998 Mar 6;92(5):587–590. doi: 10.1016/s0092-8674(00)81123-x. [DOI] [PubMed] [Google Scholar]
  44. Nakamura T. M., Cooper J. P., Cech T. R. Two modes of survival of fission yeast without telomerase. Science. 1998 Oct 16;282(5388):493–496. doi: 10.1126/science.282.5388.493. [DOI] [PubMed] [Google Scholar]
  45. Nugent C. I., Bosco G., Ross L. O., Evans S. K., Salinger A. P., Moore J. K., Haber J. E., Lundblad V. Telomere maintenance is dependent on activities required for end repair of double-strand breaks. Curr Biol. 1998 May 21;8(11):657–660. doi: 10.1016/s0960-9822(98)70253-2. [DOI] [PubMed] [Google Scholar]
  46. Nugent C. I., Hughes T. R., Lue N. F., Lundblad V. Cdc13p: a single-strand telomeric DNA-binding protein with a dual role in yeast telomere maintenance. Science. 1996 Oct 11;274(5285):249–252. doi: 10.1126/science.274.5285.249. [DOI] [PubMed] [Google Scholar]
  47. Nugent C. I., Lundblad V. The telomerase reverse transcriptase: components and regulation. Genes Dev. 1998 Apr 15;12(8):1073–1085. doi: 10.1101/gad.12.8.1073. [DOI] [PubMed] [Google Scholar]
  48. Palladino F., Laroche T., Gilson E., Pillus L., Gasser S. M. The positioning of yeast telomeres depends on SIR3, SIR4, and the integrity of the nuclear membrane. Cold Spring Harb Symp Quant Biol. 1993;58:733–746. doi: 10.1101/sqb.1993.058.01.081. [DOI] [PubMed] [Google Scholar]
  49. Polotnianka R. M., Li J., Lustig A. J. The yeast Ku heterodimer is essential for protection of the telomere against nucleolytic and recombinational activities. Curr Biol. 1998 Jul 2;8(14):831–834. doi: 10.1016/s0960-9822(98)70325-2. [DOI] [PubMed] [Google Scholar]
  50. Porter S. E., Greenwell P. W., Ritchie K. B., Petes T. D. The DNA-binding protein Hdf1p (a putative Ku homologue) is required for maintaining normal telomere length in Saccharomyces cerevisiae. Nucleic Acids Res. 1996 Feb 15;24(4):582–585. doi: 10.1093/nar/24.4.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Prescott J., Blackburn E. H. Telomerase RNA mutations in Saccharomyces cerevisiae alter telomerase action and reveal nonprocessivity in vivo and in vitro. Genes Dev. 1997 Feb 15;11(4):528–540. doi: 10.1101/gad.11.4.528. [DOI] [PubMed] [Google Scholar]
  52. Price C. M. Synthesis of the telomeric C-strand. A review. Biochemistry (Mosc) 1997 Nov;62(11):1216–1223. [PubMed] [Google Scholar]
  53. Ray A., Runge K. W. The yeast telomere length counting machinery is sensitive to sequences at the telomere-nontelomere junction. Mol Cell Biol. 1999 Jan;19(1):31–45. doi: 10.1128/mcb.19.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Reynolds A. E., Murray A. W., Szostak J. W. Roles of the 2 microns gene products in stable maintenance of the 2 microns plasmid of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Oct;7(10):3566–3573. doi: 10.1128/mcb.7.10.3566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Sandell L. L., Gottschling D. E., Zakian V. A. Transcription of a yeast telomere alleviates telomere position effect without affecting chromosome stability. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12061–12065. doi: 10.1073/pnas.91.25.12061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Sandell L. L., Zakian V. A. Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell. 1993 Nov 19;75(4):729–739. doi: 10.1016/0092-8674(93)90493-a. [DOI] [PubMed] [Google Scholar]
  57. Shampay J., Szostak J. W., Blackburn E. H. DNA sequences of telomeres maintained in yeast. Nature. 1984 Jul 12;310(5973):154–157. doi: 10.1038/310154a0. [DOI] [PubMed] [Google Scholar]
  58. Singer M. S., Gottschling D. E. TLC1: template RNA component of Saccharomyces cerevisiae telomerase. Science. 1994 Oct 21;266(5184):404–409. doi: 10.1126/science.7545955. [DOI] [PubMed] [Google Scholar]
  59. Tommerup H., Dousmanis A., de Lange T. Unusual chromatin in human telomeres. Mol Cell Biol. 1994 Sep;14(9):5777–5785. doi: 10.1128/mcb.14.9.5777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Virta-Pearlman V., Morris D. K., Lundblad V. Est1 has the properties of a single-stranded telomere end-binding protein. Genes Dev. 1996 Dec 15;10(24):3094–3104. doi: 10.1101/gad.10.24.3094. [DOI] [PubMed] [Google Scholar]
  61. Volkert F. C., Broach J. R. Site-specific recombination promotes plasmid amplification in yeast. Cell. 1986 Aug 15;46(4):541–550. doi: 10.1016/0092-8674(86)90879-2. [DOI] [PubMed] [Google Scholar]
  62. Walmsley R. M., Petes T. D. Genetic control of chromosome length in yeast. Proc Natl Acad Sci U S A. 1985 Jan;82(2):506–510. doi: 10.1073/pnas.82.2.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Watson J. D. Origin of concatemeric T7 DNA. Nat New Biol. 1972 Oct 18;239(94):197–201. doi: 10.1038/newbio239197a0. [DOI] [PubMed] [Google Scholar]
  64. Wotton D., Shore D. A novel Rap1p-interacting factor, Rif2p, cooperates with Rif1p to regulate telomere length in Saccharomyces cerevisiae. Genes Dev. 1997 Mar 15;11(6):748–760. doi: 10.1101/gad.11.6.748. [DOI] [PubMed] [Google Scholar]
  65. Wright J. H., Gottschling D. E., Zakian V. A. Saccharomyces telomeres assume a non-nucleosomal chromatin structure. Genes Dev. 1992 Feb;6(2):197–210. doi: 10.1101/gad.6.2.197. [DOI] [PubMed] [Google Scholar]
  66. Zakian V. A. Telomeres: beginning to understand the end. Science. 1995 Dec 8;270(5242):1601–1607. doi: 10.1126/science.270.5242.1601. [DOI] [PubMed] [Google Scholar]
  67. van Steensel B., Smogorzewska A., de Lange T. TRF2 protects human telomeres from end-to-end fusions. Cell. 1998 Feb 6;92(3):401–413. doi: 10.1016/s0092-8674(00)80932-0. [DOI] [PubMed] [Google Scholar]
  68. van Steensel B., de Lange T. Control of telomere length by the human telomeric protein TRF1. Nature. 1997 Feb 20;385(6618):740–743. doi: 10.1038/385740a0. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES