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

(2009) CYTOMETRY PART A. 75(5). p.428-439
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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.
Keywords
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

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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.
@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},
}

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