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. 1992 Mar 1;89(5):1695–1699. doi: 10.1073/pnas.89.5.1695

Highly conserved repetitive DNA sequences are present at human centromeres.

D L Grady 1, R L Ratliff 1, D L Robinson 1, E C McCanlies 1, J Meyne 1, R K Moyzis 1
PMCID: PMC48519  PMID: 1542662

Abstract

Highly conserved repetitive DNA sequence clones, largely consisting of (GGAAT)n repeats, have been isolated from a human recombinant repetitive DNA library by high-stringency hybridization with rodent repetitive DNA. This sequence, the predominant repetitive sequence in human satellites II and III, is similar to the essential core DNA of the Saccharomyces cerevisiae centromere, centromere DNA element (CDE) III. In situ hybridization to human telophase and Drosophila polytene chromosomes shows localization of the (GGAAT)n sequence to centromeric regions. Hyperchromicity studies indicate that the (GGAAT)n sequence exhibits unusual hydrogen bonding properties. The purine-rich strand alone has the same thermal stability as the duplex. Hyperchromicity studies of synthetic DNA variants indicate that all sequences with the composition (AATGN)n exhibit this unusual thermal stability. DNA-mobility-shift assays indicate that specific HeLa-cell nuclear proteins recognize this sequence with a relative affinity greater than 10(5). The extreme evolutionary conservation of this DNA sequence, its centromeric location, its unusual hydrogen bonding properties, its high affinity for specific nuclear proteins, and its similarity to functional centromeres isolated from yeast suggest that this sequence may be a component of the functional human centromere.

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Selected References

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

  1. Boué A., Boué J., Gropp A. Cytogenetics of pregnancy wastage. Adv Hum Genet. 1985;14:1–57. doi: 10.1007/978-1-4615-9400-0_1. [DOI] [PubMed] [Google Scholar]
  2. Carbon J., Clarke L. Centromere structure and function in budding and fission yeasts. New Biol. 1990 Jan;2(1):10–19. [PubMed] [Google Scholar]
  3. Fitzgerald-Hayes M., Clarke L., Carbon J. Nucleotide sequence comparisons and functional analysis of yeast centromere DNAs. Cell. 1982 May;29(1):235–244. doi: 10.1016/0092-8674(82)90108-8. [DOI] [PubMed] [Google Scholar]
  4. Gosden J. R., Mitchell A. R., Buckland R. A., Clayton R. P., Evans H. J. The location of four human satellite DNAs on human chromosomes. Exp Cell Res. 1975 Apr;92(1):148–158. doi: 10.1016/0014-4827(75)90648-5. [DOI] [PubMed] [Google Scholar]
  5. John B., Miklos G. L. Functional aspects of satellite DNA and heterochromatin. Int Rev Cytol. 1979;58:1–114. doi: 10.1016/s0074-7696(08)61473-4. [DOI] [PubMed] [Google Scholar]
  6. Lechner J., Carbon J. A 240 kd multisubunit protein complex, CBF3, is a major component of the budding yeast centromere. Cell. 1991 Feb 22;64(4):717–725. doi: 10.1016/0092-8674(91)90501-o. [DOI] [PubMed] [Google Scholar]
  7. Li Y., Zon G., Wilson W. D. NMR and molecular modeling evidence for a G.A mismatch base pair in a purine-rich DNA duplex. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):26–30. doi: 10.1073/pnas.88.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Li Y., Zon G., Wilson W. D. Thermodynamics of DNA duplexes with adjacent G.A mismatches. Biochemistry. 1991 Jul 30;30(30):7566–7572. doi: 10.1021/bi00244a028. [DOI] [PubMed] [Google Scholar]
  9. Lohe A. R., Brutlag D. L. Multiplicity of satellite DNA sequences in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1986 Feb;83(3):696–700. doi: 10.1073/pnas.83.3.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Maio J. J., Brown F. L., Musich P. R. Toward a molecular paleontology of primate genomes. I. The HindIII and EcoRI dimer families of alphoid DNAs. Chromosoma. 1981;83(1):103–125. doi: 10.1007/BF00286019. [DOI] [PubMed] [Google Scholar]
  11. Meyne J., Baker R. J., Hobart H. H., Hsu T. C., Ryder O. A., Ward O. G., Wiley J. E., Wurster-Hill D. H., Yates T. L., Moyzis R. K. Distribution of non-telomeric sites of the (TTAGGG)n telomeric sequence in vertebrate chromosomes. Chromosoma. 1990 Apr;99(1):3–10. doi: 10.1007/BF01737283. [DOI] [PubMed] [Google Scholar]
  12. Meyne J., Ratliff R. L., Moyzis R. K. Conservation of the human telomere sequence (TTAGGG)n among vertebrates. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7049–7053. doi: 10.1073/pnas.86.18.7049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miller D. A. Evolution of primate chromosomes. Science. 1977 Dec 16;198(4322):1116–1124. doi: 10.1126/science.929190. [DOI] [PubMed] [Google Scholar]
  14. Moyzis R. K., Albright K. L., Bartholdi M. F., Cram L. S., Deaven L. L., Hildebrand C. E., Joste N. E., Longmire J. L., Meyne J., Schwarzacher-Robinson T. Human chromosome-specific repetitive DNA sequences: novel markers for genetic analysis. Chromosoma. 1987;95(6):375–386. doi: 10.1007/BF00333988. [DOI] [PubMed] [Google Scholar]
  15. Moyzis R. K., Buckingham J. M., Cram L. S., Dani M., Deaven L. L., Jones M. D., Meyne J., Ratliff R. L., Wu J. R. 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. doi: 10.1073/pnas.85.18.6622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moyzis R. K., Torney D. C., Meyne J., Buckingham J. M., Wu J. R., Burks C., Sirotkin K. M., Goad W. B. The distribution of interspersed repetitive DNA sequences in the human genome. Genomics. 1989 Apr;4(3):273–289. doi: 10.1016/0888-7543(89)90331-5. [DOI] [PubMed] [Google Scholar]
  17. Pardue M. L., Gall J. G. Chromosomal localization of mouse satellite DNA. Science. 1970 Jun 12;168(3937):1356–1358. doi: 10.1126/science.168.3937.1356. [DOI] [PubMed] [Google Scholar]
  18. Pluta A. F., Cooke C. A., Earnshaw W. C. Structure of the human centromere at metaphase. Trends Biochem Sci. 1990 May;15(5):181–185. doi: 10.1016/0968-0004(90)90158-8. [DOI] [PubMed] [Google Scholar]
  19. Privé G. G., Heinemann U., Chandrasegaran S., Kan L. S., Kopka M. L., Dickerson R. E. Helix geometry, hydration, and G.A mismatch in a B-DNA decamer. Science. 1987 Oct 23;238(4826):498–504. doi: 10.1126/science.3310237. [DOI] [PubMed] [Google Scholar]
  20. Prosser J., Frommer M., Paul C., Vincent P. C. Sequence relationships of three human satellite DNAs. J Mol Biol. 1986 Jan 20;187(2):145–155. doi: 10.1016/0022-2836(86)90224-x. [DOI] [PubMed] [Google Scholar]
  21. Rattner J. B. The organization of the mammalian kinetochore: a scanning electron microscope study. Chromosoma. 1987;95(3):175–181. doi: 10.1007/BF00330348. [DOI] [PubMed] [Google Scholar]
  22. Rich A., RajBhandary U. L. Transfer RNA: molecular structure, sequence, and properties. Annu Rev Biochem. 1976;45:805–860. doi: 10.1146/annurev.bi.45.070176.004105. [DOI] [PubMed] [Google Scholar]
  23. Riethman H. C., Moyzis R. K., Meyne J., Burke D. T., Olson M. V. Cloning human telomeric DNA fragments into Saccharomyces cerevisiae using a yeast-artificial-chromosome vector. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6240–6244. doi: 10.1073/pnas.86.16.6240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Strauss F., Varshavsky A. A protein binds to a satellite DNA repeat at three specific sites that would be brought into mutual proximity by DNA folding in the nucleosome. Cell. 1984 Jul;37(3):889–901. doi: 10.1016/0092-8674(84)90424-0. [DOI] [PubMed] [Google Scholar]
  25. Waye J. S., Willard H. F. Human beta satellite DNA: genomic organization and sequence definition of a class of highly repetitive tandem DNA. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6250–6254. doi: 10.1073/pnas.86.16.6250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Williamson J. R., Raghuraman M. K., Cech T. R. Monovalent cation-induced structure of telomeric DNA: the G-quartet model. Cell. 1989 Dec 1;59(5):871–880. doi: 10.1016/0092-8674(89)90610-7. [DOI] [PubMed] [Google Scholar]
  27. Zinkowski R. P., Meyne J., Brinkley B. R. The centromere-kinetochore complex: a repeat subunit model. J Cell Biol. 1991 Jun;113(5):1091–1110. doi: 10.1083/jcb.113.5.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. von Hippel P. H., Berg O. G. On the specificity of DNA-protein interactions. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1608–1612. doi: 10.1073/pnas.83.6.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]

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