The architecture of interphase chromosomes and nucleolar transcription sites in plants

P.J. Shaw, R. Abranches, A.P. Santos, A.F. Beven, E. Stoger, E. Wegel, P. González-Melendi

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Fluorescence in situ hybridization (FISH) coupled with confocal microscopy has been used to reveal the interphase chromosome organization in plants. In wheat and several other related species, we have shown that the interphase chromosomes are in a very well-defined organization, with centromeres and telomeres located at opposite sides of the nuclear envelope - a classic Rabl configuration. In transgenic wheat lines, FISH analysis of metaphase chromosomes has shown that multiple transgene copies can be integrated along a single chromosome, with large regions of intervening genomic sequence. These multiple copies are often colocalized in interphase, suggesting either an ectopic association or a highly reproducible interphase chromatin configuration. Bromo-uridine (BrU) incorporation has been used to label transcription sites in the nucleolus. Using pea root tissue, we have combined BrU incorporation with preembedding 1-nm gold detection to image the nucleolar transcription sites by electron microscopy. This has revealed many distinct elongated clusters of silver-gold particles. These clusters are 200-300 nm in length and are thicker at one end than the other. We suggest that each cluster corresponds to a single transcribed gene. Serial sectioning of several entire nucleoli has enabled the reconstruction of all the nucleolar transcription sites, and we have estimated that there are 200-300 transcribed genes per nucleolus.
Original languageEnglish
Pages (from-to)31-38
Number of pages8
JournalJournal Of Structural Biology
Volume140
Issue number1-3
DOIs
Publication statusPublished - 2002

Keywords

  • rDNA, rRNA
  • Serial sectioning
  • 1 nm gold
  • 3D microscopy
  • BrU
  • v
  • Confocal microscopy
  • Electron microscopy
  • Fluorescence in situ hybridization
  • Genomic in situ hybridization
  • Interptu hyhase chromosomes
  • Nucleolus
  • Nucleus
  • Plant
  • Rabl configurationr

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    Shaw, P. J., Abranches, R., Santos, A. P., Beven, A. F., Stoger, E., Wegel, E., & González-Melendi, P. (2002). The architecture of interphase chromosomes and nucleolar transcription sites in plants. Journal Of Structural Biology, 140(1-3), 31-38. https://doi.org/10.1016/S1047-8477(02)00537-3