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

Keywords

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

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