Document Detail


Changes in chromatin compaction during the cell cycle revealed by micrometer-scale measurement of molecular flow in the nucleus.
MedLine Citation:
PMID:  22325293     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We present a quantitative fluctuation-based assay to measure the degree of local chromatin compaction and investigate how chromatin density regulates the diffusive path adopted by an inert protein in dividing cells. The assay uses CHO-K1 cells coexpressing untagged enhanced green fluorescent protein (EGFP) and histone H2B tagged mCherry. We measure at the single-cell level the EGFP localization and molecular flow patterns characteristic of each stage of chromatin compaction from mitosis through interphase by means of pair-correlation analysis. We find that the naturally occurring changes in chromatin organization impart a regulation on the spatial distribution and temporal dynamics of EGFP within the nucleus. Combined with the analysis of Ca(2+) intracellular homeostasis during cell division, EGFP flow regulation can be interpreted as the result of controlled changes in chromatin compaction. For the first time, to our knowledge, we were able to probe chromatin compaction on the micrometer scale, where the regulation of molecular diffusion may become relevant for many cellular processes.
Authors:
Elizabeth Hinde; Francesco Cardarelli; Michelle A Digman; Enrico Gratton
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-02-07
Journal Detail:
Title:  Biophysical journal     Volume:  102     ISSN:  1542-0086     ISO Abbreviation:  Biophys. J.     Publication Date:  2012 Feb 
Date Detail:
Created Date:  2012-02-13     Completed Date:  2012-06-15     Revised Date:  2013-04-11    
Medline Journal Info:
Nlm Unique ID:  0370626     Medline TA:  Biophys J     Country:  United States    
Other Details:
Languages:  eng     Pagination:  691-7     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Affiliation:
Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CHO Cells
Calcium / metabolism
Cell Cycle*
Cell Nucleus / metabolism*
Chromatin Assembly and Disassembly*
Cricetinae
Cricetulus
Diffusion
Homeostasis
Interphase
Microscopy, Fluorescence / methods*
Mitosis
Models, Molecular
Protein Conformation
Grant Support
ID/Acronym/Agency:
P41-RR03155/RR/NCRR NIH HHS; P50-GM076516/GM/NIGMS NIH HHS; U54 GM064346/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
7440-70-2/Calcium
Comments/Corrections

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