Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope
Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon micros...
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| Published in | Biophysical journal Vol. 89; no. 6; pp. 4275 - 4285 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Elsevier Inc
01.12.2005
Biophysical Society |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon microscope that provides ∼10-fold higher spatial and temporal resolutions than previous measurements. We followed the motion of a chromatin sequence containing a lac-operator repeat in cells stably expressing lac repressor fused with enhanced green fluorescent protein, observing long periods of apparent constrained diffusion interrupted by relatively abrupt jumps of ∼150
nm lasting 0.3–2
s. During these jumps, the particle moved an average of four times faster than in the periods between jumps and in paths more rectilinear than predicted for random diffusion motion. Additionally, the jumps were sensitive to the temperature and absent after ATP depletion. These experimental results point to an energy-dependent mechanism driving fast motion of chromatin in interphase cells. |
|---|---|
| AbstractList | Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon microscope that provides ∼10-fold higher spatial and temporal resolutions than previous measurements. We followed the motion of a chromatin sequence containing a lac-operator repeat in cells stably expressing lac repressor fused with enhanced green fluorescent protein, observing long periods of apparent constrained diffusion interrupted by relatively abrupt jumps of ∼150
nm lasting 0.3–2
s. During these jumps, the particle moved an average of four times faster than in the periods between jumps and in paths more rectilinear than predicted for random diffusion motion. Additionally, the jumps were sensitive to the temperature and absent after ATP depletion. These experimental results point to an energy-dependent mechanism driving fast motion of chromatin in interphase cells. Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon microscope that provides ∼10-fold higher spatial and temporal resolutions than previous measurements. We followed the motion of a chromatin sequence containing a lac-operator repeat in cells stably expressing lac repressor fused with enhanced green fluorescent protein, observing long periods of apparent constrained diffusion interrupted by relatively abrupt jumps of ∼150 nm lasting 0.3–2 s. During these jumps, the particle moved an average of four times faster than in the periods between jumps and in paths more rectilinear than predicted for random diffusion motion. Additionally, the jumps were sensitive to the temperature and absent after ATP depletion. These experimental results point to an energy-dependent mechanism driving fast motion of chromatin in interphase cells. Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon microscope that provides approximately 10-fold higher spatial and temporal resolutions than previous measurements. We followed the motion of a chromatin sequence containing a lac-operator repeat in cells stably expressing lac repressor fused with enhanced green fluorescent protein, observing long periods of apparent constrained diffusion interrupted by relatively abrupt jumps of approximately 150 nm lasting 0.3-2 s. During these jumps, the particle moved an average of four times faster than in the periods between jumps and in paths more rectilinear than predicted for random diffusion motion. Additionally, the jumps were sensitive to the temperature and absent after ATP depletion. These experimental results point to an energy-dependent mechanism driving fast motion of chromatin in interphase cells. Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned within nuclei remain unclear. Here, we study the dynamics of chromatin in vivo applying a novel particle-tracking method in a two-photon microscope that provides ~10-fold higher spatial and temporal resolutions than previous measurements. We followed the motion of a chromatin sequence containing a lac-operator repeat in cells stably expressing lac repressor fused with enhanced green fluorescent protein, observing long periods of apparent constrained diffusion interrupted by relatively abrupt jumps of ~150 nm lasting 0.3-2 s. During these jumps, the particle moved an average of four times faster than in the periods between jumps and in paths more rectilinear than predicted for random diffusion motion. Additionally, the jumps were sensitive to the temperature and absent after ATP depletion. These experimental results point to an energy-dependent mechanism driving fast motion of chromatin in interphase cells. [PUBLICATION ABSTRACT] |
| Author | Levi, Valeria Belmont, Andrew S. Ruan, QiaoQiao Gratton, Enrico Plutz, Matthew |
| AuthorAffiliation | Laboratory for Fluorescence Dynamics, and † Department of Cell and Structural Biology, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois |
| AuthorAffiliation_xml | – name: Laboratory for Fluorescence Dynamics, and † Department of Cell and Structural Biology, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois |
| Author_xml | – sequence: 1 givenname: Valeria surname: Levi fullname: Levi, Valeria organization: Laboratory for Fluorescence Dynamics, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois – sequence: 2 givenname: QiaoQiao surname: Ruan fullname: Ruan, QiaoQiao organization: Laboratory for Fluorescence Dynamics, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois – sequence: 3 givenname: Matthew surname: Plutz fullname: Plutz, Matthew organization: Department of Cell and Structural Biology, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois – sequence: 4 givenname: Andrew S. surname: Belmont fullname: Belmont, Andrew S. organization: Department of Cell and Structural Biology, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois – sequence: 5 givenname: Enrico surname: Gratton fullname: Gratton, Enrico email: enrico@scs.uiuc.edu organization: Laboratory for Fluorescence Dynamics, Chemical and Life Science Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/16150965$$D View this record in MEDLINE/PubMed |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Address reprint requests to Enrico Gratton, Laboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, 1110 West Green St., Urbana, IL 61801-3080. Tel.: 217-244-5620; Fax: 217-244-7187; E-mail: enrico@scs.uiuc.edu. |
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| PublicationYear | 2005 |
| Publisher | Elsevier Inc Biophysical Society |
| Publisher_xml | – name: Elsevier Inc – name: Biophysical Society |
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| Snippet | Increasing evidence points to a dynamical compartmentalization of the cell nucleus, yet the mechanisms by which interphase chromatin moves and is positioned... |
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| SubjectTerms | Animals Cell Nucleus - physiology Cells Changes CHO Cells Chromatin Chromatin - physiology Chromatin - ultrastructure Cricetinae Cricetulus Diffusion Digital imaging Dihydrofolate reductase Genes Interphase - physiology Life sciences Methods Microscopy Microscopy, Fluorescence, Multiphoton - methods Molecular biology Monoclonal antibodies Motion Proteins RNA polymerase Spectroscopy, Imaging, Other Techniques |
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| Title | Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope |
| URI | https://dx.doi.org/10.1529/biophysj.105.066670 https://www.ncbi.nlm.nih.gov/pubmed/16150965 https://www.proquest.com/docview/215700083 https://search.proquest.com/docview/68843571 https://pubmed.ncbi.nlm.nih.gov/PMC1366992 |
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