Correlative single molecule lattice light sheet imaging reveals the dynamic relationship between nucleosomes and the local chromatin environment.
Timothy A DaugirdYu ShiKatie L HollandHosein RostamianZhe J LiuLuke D LavisJoseph RodriguezBrian D StrahlWesley R LegantPublished in: Nature communications (2024)
In the nucleus, biological processes are driven by proteins that diffuse through and bind to a meshwork of nucleic acid polymers. To better understand this interplay, we present an imaging platform to simultaneously visualize single protein dynamics together with the local chromatin environment in live cells. Together with super-resolution imaging, new fluorescent probes, and biophysical modeling, we demonstrate that nucleosomes display differential diffusion and packing arrangements as chromatin density increases whereas the viscoelastic properties and accessibility of the interchromatin space remain constant. Perturbing nuclear functions impacts nucleosome diffusive properties in a manner that is dependent both on local chromatin density and on relative location within the nucleus. Our results support a model wherein transcription locally stabilizes nucleosomes while simultaneously allowing for the free exchange of nuclear proteins. Additionally, they reveal that nuclear heterogeneity arises from both active and passive processes and highlight the need to account for different organizational principles when modeling different chromatin environments.
Keyphrases
- single molecule
- transcription factor
- genome wide
- dna damage
- gene expression
- high resolution
- nucleic acid
- living cells
- atomic force microscopy
- induced apoptosis
- dna methylation
- fluorescence imaging
- single cell
- small molecule
- high throughput
- oxidative stress
- quantum dots
- mass spectrometry
- cell death
- signaling pathway
- high speed
- protein protein