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Controlled light exposure microscopy reveals dynamic telomere microterritories throughout the cell cycle

Winnok De Vos UGent, Ron A Hoebe, Greg H Joss, Willem Haffmans, Sarah Baatout, Patric Van Oostveldt UGent and Erik MM Manders (2009) CYTOMETRY PART A. 75(5). p.428-439
abstract
Telomeres are complex end structures that confer functional integrity and positional stability to human chromosomes. Despite their critical importance, there is no clear view on telomere organization in cycling human cells and their dynamic behavior throughout the cell cycle. We investigated spatiotemporal organization of telomeres in living human ECV-304 cells stably expressing telomere binding proteins TRF1 and TRF2 fused to mCitrine using four dimensional microscopy. We thereby made use of controlled light exposure microscopy (CLEM), a novel technology that strongly reduces photodamage by limiting excitation in parts of the image where full exposure is not needed. We found that telomeres share small territories where they dynamically associate. These territories are preferentially positioned at the interface of chromatin domains. TRF1 and TRF2 are abundantly present in these territories but not firmly bound. At the onset of mitosis, the bulk of TRF protein dissociates from telomere regions, territories disintegrate and individual telomeres become faintly visible. The combination of stable cell lines, CLEM and cytometry proved essential in providing novel insights in compartment-based nuclear organization and may serve as a model approach for investigating telomere-driven genome-instability and studying long-term nuclear dynamics.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
HUMAN-LYMPHOCYTES, PROTEIN TRF2, nuclear organization, TRF2, chromatin dynamics, live cell imaging, CLEM, PHOTOBLEACHING KINETICS, LIVING MAMMALIAN-CELLS, FLUORESCENCE MICROSCOPY, INTERPHASE NUCLEUS, DNA-DAMAGE, NUCLEAR ARCHITECTURE, CONFOCAL MICROSCOPY, telomere, ENDOTHELIAL-CELLS, TRF1
journal title
CYTOMETRY PART A
Cytom. Part A
volume
75
issue
5
pages
428 - 439
Web of Science type
Article
Web of Science id
000265446300007
JCR category
BIOCHEMICAL RESEARCH METHODS
JCR impact factor
3.032 (2009)
JCR rank
21/65 (2009)
JCR quartile
2 (2009)
ISSN
1552-4922
DOI
10.1002/cyto.a.20699
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
663143
handle
http://hdl.handle.net/1854/LU-663143
date created
2009-05-21 11:07:31
date last changed
2016-12-19 15:43:35
@article{663143,
  abstract     = {Telomeres are complex end structures that confer functional integrity and positional stability to human chromosomes. Despite their critical importance, there is no clear view on telomere organization in cycling human cells and their dynamic behavior throughout the cell cycle. We investigated spatiotemporal organization of telomeres in living human ECV-304 cells stably expressing telomere binding proteins TRF1 and TRF2 fused to mCitrine using four dimensional microscopy. We thereby made use of controlled light exposure microscopy (CLEM), a novel technology that strongly reduces photodamage by limiting excitation in parts of the image where full exposure is not needed. We found that telomeres share small territories where they dynamically associate. These territories are preferentially positioned at the interface of chromatin domains. TRF1 and TRF2 are abundantly present in these territories but not firmly bound. At the onset of mitosis, the bulk of TRF protein dissociates from telomere regions, territories disintegrate and individual telomeres become faintly visible. The combination of stable cell lines, CLEM and cytometry proved essential in providing novel insights in compartment-based nuclear organization and may serve as a model approach for investigating telomere-driven genome-instability and studying long-term nuclear dynamics.},
  author       = {De Vos, Winnok and Hoebe, Ron A and Joss, Greg H and Haffmans, Willem and Baatout, Sarah and Van Oostveldt, Patric and Manders, Erik MM},
  issn         = {1552-4922},
  journal      = {CYTOMETRY PART A},
  keyword      = {HUMAN-LYMPHOCYTES,PROTEIN TRF2,nuclear organization,TRF2,chromatin dynamics,live cell imaging,CLEM,PHOTOBLEACHING KINETICS,LIVING MAMMALIAN-CELLS,FLUORESCENCE MICROSCOPY,INTERPHASE NUCLEUS,DNA-DAMAGE,NUCLEAR ARCHITECTURE,CONFOCAL MICROSCOPY,telomere,ENDOTHELIAL-CELLS,TRF1},
  language     = {eng},
  number       = {5},
  pages        = {428--439},
  title        = {Controlled light exposure microscopy reveals dynamic telomere microterritories throughout the cell cycle},
  url          = {http://dx.doi.org/10.1002/cyto.a.20699},
  volume       = {75},
  year         = {2009},
}

Chicago
De Vos, Winnok, Ron A Hoebe, Greg H Joss, Willem Haffmans, Sarah Baatout, Patric Van Oostveldt, and Erik MM Manders. 2009. “Controlled Light Exposure Microscopy Reveals Dynamic Telomere Microterritories Throughout the Cell Cycle.” Cytometry Part A 75 (5): 428–439.
APA
De Vos, Winnok, Hoebe, R. A., Joss, G. H., Haffmans, W., Baatout, S., Van Oostveldt, P., & Manders, E. M. (2009). Controlled light exposure microscopy reveals dynamic telomere microterritories throughout the cell cycle. CYTOMETRY PART A, 75(5), 428–439.
Vancouver
1.
De Vos W, Hoebe RA, Joss GH, Haffmans W, Baatout S, Van Oostveldt P, et al. Controlled light exposure microscopy reveals dynamic telomere microterritories throughout the cell cycle. CYTOMETRY PART A. 2009;75(5):428–39.
MLA
De Vos, Winnok, Ron A Hoebe, Greg H Joss, et al. “Controlled Light Exposure Microscopy Reveals Dynamic Telomere Microterritories Throughout the Cell Cycle.” CYTOMETRY PART A 75.5 (2009): 428–439. Print.