About: Fine structure of the %22PcG body%22 in human U-2 OS cells established by correlative light-electron microscopy     Goto   Sponge   NotDistinct   Permalink

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Description
  • Polycomb group (PcG) proteins of the Polycomb repressive complex 1 (PRC1) are found to be diffusely distributed in nuclei of cells from various species. However they can also be localized in intensely fluorescent foci, whether imaged using GFP fusions to proteins of PRC1 complex, or by conventional immunofluorescence microscopy. Such foci are termed Polycomb bodies (PcG bodies), and are believed to be situated into the nuclear intechromatin compartment. However, an ultrastructural description of the PcG body has not been reported to date. To establish the ultrastructure of PcG bodies in human U-2 OS cells stably expressing recombinant polycomb BMI1-GFP protein, we used correlative light electron microscopy (CLEM) implemented with high-pressure freezing, cryosubstitution and on section labeling of BMI1 protein with immunogold. This approach allowed us to clearly identify fluorescent PcG bodies, not as distinct nuclear bodies, but as nuclear domains enriched in separated heterochromatin fascicles. Importantly, high-pressure freezing and cryosubstitution allowed for a high and clear-cut imunogold BMI1 labeling of heterochromatin structures throughout the nucleus. The density of immunogold labeled BMI1 in the heterochromatin fascicles corresponding to fluorescent %22PcG bodies%22 did not differ from the density of labeling of heterochromatin fascicles outside of the %22PcG bodies.%22 Accordingly, an appearance of the fluorescent %22PcG bodies%22 seems to reflect a local accumulation of the labeled heterochromatin structures in the investigated cells. The results of this study should allow to expand the knowledge about the biological relevance of the %22PcG bodies%22 in human cells.
  • Polycomb group (PcG) proteins of the Polycomb repressive complex 1 (PRC1) are found to be diffusely distributed in nuclei of cells from various species. However they can also be localized in intensely fluorescent foci, whether imaged using GFP fusions to proteins of PRC1 complex, or by conventional immunofluorescence microscopy. Such foci are termed Polycomb bodies (PcG bodies), and are believed to be situated into the nuclear intechromatin compartment. However, an ultrastructural description of the PcG body has not been reported to date. To establish the ultrastructure of PcG bodies in human U-2 OS cells stably expressing recombinant polycomb BMI1-GFP protein, we used correlative light electron microscopy (CLEM) implemented with high-pressure freezing, cryosubstitution and on section labeling of BMI1 protein with immunogold. This approach allowed us to clearly identify fluorescent PcG bodies, not as distinct nuclear bodies, but as nuclear domains enriched in separated heterochromatin fascicles. Importantly, high-pressure freezing and cryosubstitution allowed for a high and clear-cut imunogold BMI1 labeling of heterochromatin structures throughout the nucleus. The density of immunogold labeled BMI1 in the heterochromatin fascicles corresponding to fluorescent %22PcG bodies%22 did not differ from the density of labeling of heterochromatin fascicles outside of the %22PcG bodies.%22 Accordingly, an appearance of the fluorescent %22PcG bodies%22 seems to reflect a local accumulation of the labeled heterochromatin structures in the investigated cells. The results of this study should allow to expand the knowledge about the biological relevance of the %22PcG bodies%22 in human cells. (en)
Title
  • Fine structure of the %22PcG body%22 in human U-2 OS cells established by correlative light-electron microscopy
  • Fine structure of the %22PcG body%22 in human U-2 OS cells established by correlative light-electron microscopy (en)
skos:prefLabel
  • Fine structure of the %22PcG body%22 in human U-2 OS cells established by correlative light-electron microscopy
  • Fine structure of the %22PcG body%22 in human U-2 OS cells established by correlative light-electron microscopy (en)
skos:notation
  • RIV/00216208:11110/11:10482!RIV12-MSM-11110___
http://linked.open...avai/predkladatel
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(LC535), Z(MSM0021620806)
http://linked.open...iv/cisloPeriodika
  • 3
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 199774
http://linked.open...ai/riv/idVysledku
  • RIV/00216208:11110/11:10482
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Polycomb proteins; Polycomb body; BMI1 protein; heterochromatin; correlative light-electron microscopy; high-pressure freezing; immunogold labeling (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • US - Spojené státy americké
http://linked.open...ontrolniKodProRIV
  • [4429B01DA726]
http://linked.open...i/riv/nazevZdroje
  • Nucleus
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 2
http://linked.open...iv/tvurceVysledku
  • Raška, Ivan
  • Jůda, Pavel
  • Cmarko, Dušan
  • Šmigová, Jana
http://linked.open...n/vavai/riv/zamer
issn
  • 1949-1034
number of pages
http://localhost/t...ganizacniJednotka
  • 11110
is http://linked.open...avai/riv/vysledek of
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