Detail of Fig2b_DNAunderwind
Project
Title
Z-series Lattice-SIM images of DAPI, bio-psoralen and RNAPIIS2P in nucleus
Description
Z-series Lattice-SIM images of DAPI, bio-psoralen and RNAPIIS2P in nucleus. The underwound DNA was labeled with biotinylated psoralen, which preferentially binds to underwound DNA rather than relaxed DNA, before cell fixation. Channel1;RNAPIIS2P (RNA polymerase II phosphorylated at serine 2), Chennel2;DAPI, Channel3;psoralen
Release, Updated
2025-01-31
License
CC BY
Kind
Image data
File Formats
.czi
Data size
840.3 MB
Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
nucleus
Biological Imaging Method
Lattice-pattern structured illumination microscopy
X scale
0.05 micrometer
Y scale
0.05 micrometer
Z scale
0.40 micrometer
T scale
-
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
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Objective model
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Filter set
-
Summary of Methods
Fukute J, Maki K, Adachi T. The nucleolar shell provides anchoring sites for DNA untwisting. Commun Biol. 2024 Jan 23;7(1):83.
Related paper(s)
Jumpei Fukute, Koichiro Maki, Taiji Adachi (2024) The nucleolar shell provides anchoring sites for DNA untwisting., Communications biology, Volume 7, Number 1, pp. 83
Published in 2024 Jan 23 (Electronic publication in Jan. 23, 2024, midnight )
- doi: 10.1038/s42003-023-05750-w
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pmid: 38263258
(Abstract) DNA underwinding (untwisting) is a crucial step in transcriptional activation. DNA underwinding occurs between the site where torque is generated by RNA polymerase (RNAP) and the site where the axial rotation of DNA is constrained. However, what constrains DNA axial rotation in the nucleus is yet unknown. Here, we show that the anchorage to the nuclear protein condensates constrains DNA axial rotation for DNA underwinding in the nucleolus. In situ super-resolution imaging of underwound DNA reveal that underwound DNA accumulates in the nucleolus, a nuclear condensate with a core-shell structure. Specifically, underwound DNA is distributed in the nucleolar core owing to RNA polymerase I (RNAPI) activities. Furthermore, underwound DNA in the core decreases when nucleolar shell components are prevented from binding to their recognition structure, G-quadruplex (G4). Taken together, these results suggest that the nucleolar shell provides anchoring sites that constrain DNA axial rotation for RNAPI-driven DNA underwinding in the core. Our findings will contribute to understanding how nuclear protein condensates make up constraints for the site-specific regulation of DNA underwinding and transcription.(MeSH Terms)
Contact
Koichiro Maki
, Kyoto University
, Laboratory of Cellular and Molecular Biomechanics, Department of Mammalian Regulatory Network, Graduate School of Biostudies
, Laboratory of Cellular and Molecular Biomechanics, Department of Mammalian Regulatory Network, Graduate School of Biostudies
Contributors
Jumpei Fukute, Koichiro Maki, Taiji Adachi
OMERO Project
Source