CHROMOSOMES: Major Discoveries/Highlights



Centromeric DNA: Sequences and structure in chromosomes and nuclei:

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  • Identification, isolation, sequencing and localization of human centromeric DNA repeats (1-4) with development of high-resolution in-situ hybridization using biotin-labeled nucleotides (5)

  • Discovery of tissue-specific arrangements of centromeres in interphase nuclei (6)

  • Uncovered 3D movements of centromeres during differentiation & stress in neurons (7,8)

  • Showed species-specific DNA sequences maintain a conserved tissue-specific arrangement of centromeres in interphase neuron and glial nuclei during mammalian evolution (9)

  • Glioblastomas (brain tumors) lose tissue-specific specific centromere arrangements




Discovery of human Long interspersed Repeats (LINES) and their organization

  • Identification, isolation and sequencing of LINES, including ORFs, and murine homologies (10-12)

  • LINES collect in bands on chromosome arms, shown by non-isotopic in-situ hybridization (12)

  • LINES cluster with tissue specific genes in Giemsa-dark chromosome bands whereas short ALU elements (SINES) concentrate with early replicating housekeeping genes in Giemsa-light bands, as shown by high resolution in-situ hybridization and pulse-field DNA analysis (13,14)

  • Endogenous retroviruses have species-specific addresses on chromosome arms (15)

  • Whole chromosomes are maintained as cohesive compact structures in interphase nuclei, and not in diffusely extended spaghetti-like strands (16). Small regions open locally during transcription and replication.

  • Application of in-situ detection of interphase chromosomes for tumors and genetic diseases (17,18)

  • Genetic instability is a key feature of glioblastomas (19)


A unified model of chromosome structure in interphase and metaphase

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  • Unified model for chromosome DNA compaction in heterochromatin and local unfolding of chromatids for organized transcription and replication of long (megabase) genomic DNA (20-22)

  • Transgenic insertion of tandem ORF DNA arrays into chromosome arms leads to gene silencing with heterochromatic ultrastructure, and its positioning with centromere domains in nuclei (23)





SUMMARY

Identified DNA repeats are not “junk”, but are critical recognition elements that define specific chromosome regions and structures, and contribute to the function of organized gene sets.


Genetic instability in glioblastomas is a key challenge for targeted treatments.


REFERENCES (SEE BELOW for MORE on DNA-CHROMOSOME)

  1. Manuelidis, L., Repeating restriction fragments of human DNA. Nucleic Acids Res. 3:3063-3076, 1976.
  2. Manuelidis, L., Complex and simple sequences in human repeated DNAs. Chromosoma 66:1-21, 1978.
  3. Manuelidis, L., Chromosomal localization of complex and simple repeated human DNAs. Chromosoma 66:23-32, 1978.
  4. Wu, J.C. and Manuelidis, L., Sequence definition and organization of a human repeated DNA. J. Mol. Biol. 142:363-386, 1980.
  5. Manuelidis, L., Langer-Safer, P.R. and Ward, D.C., High resolution mapping of satellite DNA using biotin-labeled DNA probes. J. Cell Biol. 95:619-625, 1982.
  6. Manuelidis, L. and Borden, J., Reproducible compartmentalization of individual chromosome domains in human CNS cells revealed by in situ hybridization and three-dimensional reconstruction. Chromosoma 96:397-401, 1988.
  7. Manuelidis, L., Indications of centromere movement during interphase and differentiation. Ann. N.Y. Acad. Sci. 450:205-221, 1985.
  8. Borden, J. and Manuelidis, L., Movement of the X chromosome in epilepsy. Science 242:1687-1691, 1988.
  9. Manuelidis, L., Different CNS cell types display distinct and non-random arrangements of satellite DNA sequences, Proc. Nat. Acad. Sci. USA 181:3123-3127, 1984.
  10. Manuelidis, L., Novel classes of mouse repeated DNAs. Nucleic Acids Res. 8:3247-3258, 1980.
  11. Manuelidis, L., Nucleotide sequence definition of a major human DNA, the Hind III 1.9 kb family. Nucleic Acids Res. 10:3211-3219, 1982.
  12. Manuelidis, L., Repeated DNA sequences and nuclear structure. In: Genome Evolution (Dover, G.A. and Flavell, R.B., eds.), Academic Press, Inc., pp. 263-285, 1982.
  13. Manuelidis, L. and Ward, D.C., Chromosomal and nuclear distribution of the Hind III 1.9 kb repeat segment. Chromosoma 91:28-38, 1984.
  14. Chen, T.L. and Manuelidis, L., SINEs and LINEs cluster in distinct DNA fragments of Giemsa band size. Chromosoma 98:309-316, 1989.
  15. Taruscio, D., Manuelidis, L., Integration site preferences of endogenous retroviruses. Chromosoma 101:141-156, 1991
  16. Manuelidis, L., Individual interphase chromosome domains revealed by in situ hybridization. Human Genet. 71:288-293, 1985.
  17. Cremer, T., Lichter, P., Borden, J., Ward, D.C. and Manuelidis, L., Detection of chromosome aberrations in interphase tumor cells by in situ hybridization using chromosome specific library probes. Hum. Genet. 80:235-246, 1988.
  18. Lichter, P., Cremer, T., Tang, C., Watkins, P.C., Manuelidis, L. and Ward, D.C., Rapid detection of human chromosome 21 aberrations by in situ hybridization. Proc. Natl. Acad. Sci. USA 85:9664-9668, 1988.
  19. Manuelidis, L., Genomic stability and instability in different neuroepithelial tumors: A role for chromosome structure? J. Neuro-Oncol, 18:225-239, 1994.
  20. Sedat, J. and Manuelidis, L., A direct approach to the structure of eukaryotic chromosomes. Cold Spring Harbor Symp. Quant. Biol. 42:331-350, 1978.
  21. Manuelidis, L. and Chen, T.L., A unified model of eukaryotic chromosomes. Cytometry 18:8-25, 1990.
  22. Manuelidis, L., A view of interphase chromosomes. Science 250:1533-1540, 1990.
  23. Manuelidis, L., Heterochromatic features of an 11 Mb transgene in brain cells. Proc. Natl. Acad. Sci. USA 88:1049-1053,
Model Chromosome

Chromosome/DNA structure



A view of interphase chromosomes


Manuelidis L.
Science 1990, 250:1533-1540
Abstract
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A unified model of eukaryotic chromosomes


Manuelidis L, Chen T.
Cytometry 1990, 11:8-25
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Genomic stability and instability in different neuroepithelial tumors. A role for chromosome structure?

Manuelidis L.
Journal of Neuro-Oncology 1994, 18:225-239
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Interphase chromosome positions and structure during silencing, transcription and replication


Manuelidis L.
From: Nuclear organization, chromatin structure and gene expression. Roel Van Driel & Arie P. Otte, eds. 1997.
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