Quantitative FLIM-FRET Microscopy to Monitor Nanoscale Chromatin Compaction In Vivo Reveals Structural Roles of Condensin Complexes

David Llères (Lead / Corresponding author), Aymeric P. Bailly, Aurélien Perrin, David G. Norman, Dimitris P. Xirodimas, Robert Feil

Research output: Contribution to journalArticle

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Abstract

How metazoan genomes are structured at the nanoscale in living cells and tissues remains unknown. Here, we adapted a quantitative FRET (Förster resonance energy transfer)-based fluorescence lifetime imaging microscopy (FLIM) approach to assay nanoscale chromatin compaction in living organisms. Caenorhabditis elegans was chosen as a model system. By measuring FRET between histone-tagged fluorescent proteins, we visualized distinct chromosomal regions and quantified the different levels of nanoscale compaction in meiotic cells. Using RNAi and repetitive extrachromosomal array approaches, we defined the heterochromatin state and showed that its architecture presents a nanoscale-compacted organization controlled by Heterochromatin Protein-1 (HP1) and SETDB1 H3-lysine-9 methyltransferase homologs in vivo. Next, we functionally explored condensin complexes. We found that condensin I and condensin II are essential for heterochromatin compaction and that condensin I additionally controls lowly compacted regions. Our data show that, in living animals, nanoscale chromatin compaction is controlled not only by histone modifiers and readers but also by condensin complexes.

Original languageEnglish
Pages (from-to)1791-1803
Number of pages13
JournalCell Reports
Volume18
Issue number7
DOIs
Publication statusPublished - 14 Feb 2017

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Optical Imaging
Energy Transfer
Energy transfer
Chromatin
Microscopy
Microscopic examination
Compaction
Fluorescence
Imaging techniques
Heterochromatin
Histones
Caenorhabditis elegans
Methyltransferases
RNA Interference
Lysine
Assays
Animals
Genes
Cells
condensin complexes

Keywords

  • Journal Article
  • FLIM-FRET imaging
  • C. elegans
  • Chromatin compaction
  • Heterochromatin
  • condensin
  • HP1
  • Meiosis
  • Chromosome structure

Cite this

Llères, David ; Bailly, Aymeric P. ; Perrin, Aurélien ; Norman, David G. ; Xirodimas, Dimitris P. ; Feil, Robert. / Quantitative FLIM-FRET Microscopy to Monitor Nanoscale Chromatin Compaction In Vivo Reveals Structural Roles of Condensin Complexes. In: Cell Reports. 2017 ; Vol. 18, No. 7. pp. 1791-1803.
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Quantitative FLIM-FRET Microscopy to Monitor Nanoscale Chromatin Compaction In Vivo Reveals Structural Roles of Condensin Complexes. / Llères, David (Lead / Corresponding author); Bailly, Aymeric P.; Perrin, Aurélien; Norman, David G.; Xirodimas, Dimitris P.; Feil, Robert.

In: Cell Reports, Vol. 18, No. 7, 14.02.2017, p. 1791-1803.

Research output: Contribution to journalArticle

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AU - Llères, David

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AU - Perrin, Aurélien

AU - Norman, David G.

AU - Xirodimas, Dimitris P.

AU - Feil, Robert

N1 - D.L. was supported by a Cancéropole GSO-Emergence grant ( 2014-E17 ). R.F. acknowledges grant support from the Fondation Recherche Médicale (FRM; grant DEQ20150331703 ) and the Agence Nationale de Recherche (ANR; grant “IMPRINT-RNA” ). A.P.B. is supported by the Agence Nationale de la Recherche (ANR; grant “CeLeNeD” ), the FRM (grant SPF20120523917 ), and INSERM.

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N2 - How metazoan genomes are structured at the nanoscale in living cells and tissues remains unknown. Here, we adapted a quantitative FRET (Förster resonance energy transfer)-based fluorescence lifetime imaging microscopy (FLIM) approach to assay nanoscale chromatin compaction in living organisms. Caenorhabditis elegans was chosen as a model system. By measuring FRET between histone-tagged fluorescent proteins, we visualized distinct chromosomal regions and quantified the different levels of nanoscale compaction in meiotic cells. Using RNAi and repetitive extrachromosomal array approaches, we defined the heterochromatin state and showed that its architecture presents a nanoscale-compacted organization controlled by Heterochromatin Protein-1 (HP1) and SETDB1 H3-lysine-9 methyltransferase homologs in vivo. Next, we functionally explored condensin complexes. We found that condensin I and condensin II are essential for heterochromatin compaction and that condensin I additionally controls lowly compacted regions. Our data show that, in living animals, nanoscale chromatin compaction is controlled not only by histone modifiers and readers but also by condensin complexes.

AB - How metazoan genomes are structured at the nanoscale in living cells and tissues remains unknown. Here, we adapted a quantitative FRET (Förster resonance energy transfer)-based fluorescence lifetime imaging microscopy (FLIM) approach to assay nanoscale chromatin compaction in living organisms. Caenorhabditis elegans was chosen as a model system. By measuring FRET between histone-tagged fluorescent proteins, we visualized distinct chromosomal regions and quantified the different levels of nanoscale compaction in meiotic cells. Using RNAi and repetitive extrachromosomal array approaches, we defined the heterochromatin state and showed that its architecture presents a nanoscale-compacted organization controlled by Heterochromatin Protein-1 (HP1) and SETDB1 H3-lysine-9 methyltransferase homologs in vivo. Next, we functionally explored condensin complexes. We found that condensin I and condensin II are essential for heterochromatin compaction and that condensin I additionally controls lowly compacted regions. Our data show that, in living animals, nanoscale chromatin compaction is controlled not only by histone modifiers and readers but also by condensin complexes.

KW - Journal Article

KW - FLIM-FRET imaging

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KW - Heterochromatin

KW - condensin

KW - HP1

KW - Meiosis

KW - Chromosome structure

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