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Sci Rep. 2014 Mar 26;4:4477. doi: 10.1038/srep04477.

Super-resolution microscopy reveals decondensed chromatin structure at transcription sites.

Author information

1
Mechanobiology Institute, Singapore.
2
1] Laboratory of Atomic and Solid State Physics, Department of Physics [2] Howard Hughes Medical Institute Cornell University, Ithaca, New York 14853, USA.
3
1] Mechanobiology Institute, Singapore [2] Department of Biological Sciences, National University of Singapore, Singapore.

Abstract

Remodeling of the local chromatin structure is essential for the regulation of gene expression. While a number of biochemical and bioimaging experiments suggest decondensed chromatin structures are associated with transcription, a direct visualization of DNA and transcriptionally active RNA polymerase II (RNA pol II) at super-resolution is still lacking. Here we investigate the structure of chromatin isolated from HeLa cells using binding activatable localization microscopy (BALM). The sample preparation method preserved the structural integrity of chromatin. Interestingly, BALM imaging of the chromatin spreads revealed the presence of decondensed chromatin as gap structures along the spreads. These gaps were enriched with phosphorylated S5 RNA pol II, and were sensitive to the cellular transcriptional state. Taken together, we could visualize the decondensed chromatin regions together with active RNA pol II for the first time using super-resolution microscopy.

PMID:
24667378
PMCID:
PMC3966049
DOI:
10.1038/srep04477
[Indexed for MEDLINE]
Free PMC Article

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