Cell Cycle-Dependent Distribution of Telomeres, Centromeres, and Chromosome-Specific Subsatellite Domains in the Interphase Nucleus of Mouse Lymphocytes

Vourc’h, Claire; Taruscio, Domenica; Boyle, Ann; Ward, David C.

doi:10.1006/excr.1993.1068

Cited by 154 publications

(92 citation statements)

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“…An increasing body of evidence shows that telomeres are organized in a nonrandom, cell-type, and cell cycle-dependent manner (16,20,21,44,45). Although yeast cells are known to form few large clusters at the heterochromatic nuclear border (9,10), most human telomeres have been shown to avoid the nuclear periphery (16,21).…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2008

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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. ' 2008 International Society for Advancement of Cytometry Key termstelomere; TRF1; TRF2; nuclear organization; chromatin dynamics; CLEM; live cell imaging TELOMERES are the natural ends of linear chromosomes. A mammalian telomere consists of a double stranded array of simple TTAGGG repeats ending in a single stranded overhang that folds back to form a T-loop structure (1). In combination with sufficient telomere repeats, a complex of indirect and direct telomere binding proteins, dubbed shelterin, assures proper telomere structure and function. The specificity of shelterin for telomeric DNA is due to the recognition of TTAGGG repeats by three of its components: the homodimers telomeric repeat binding factor 1 and 2 (TRF1 and TRF2) that bind the duplex part of telomeres, and protection of telomeres 1 that binds to the single stranded TTAGGG repeats present at the three-overhang and in the D loop of the T-loop configuration (2-5).The nucleus has a meticulous organization with functional relevance to the cell assuring proper gene expression, replication and genome stability (6-8). Telomeres are considered to play an important role in spatial genome organization. Whereas yeast telomeres are known to form few large silencing clusters at the heterochromatic nuclear periphery (9,10), there is no consensus on how telomeres are organized in the mammalian nucleus. From an architectural point of view, human telomeres have been proposed to be anchored to a nuclear scaffold, thereby possibly providing struc-

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Section: Discussionmentioning

confidence: 99%

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

et al. 2008

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“…This is an open question, since the available data do not allow for a consensus in interpretation. While some research groups do not find substantial chromosomal movements [Abney et al, 1997;Gerlich et al, 2003], others find chromosomal reorganization during the cell cycle [Ferguson and Ward, 1992;Vourc'h et al, 1993;Bridger et al, 2000;Chubb et al, 2002;Walter et al, 2003;Essers et al, 2005], cellular differentiation [Stadler et al, 2004], and during quiescence and senescence [Bridger et al, 2000].…”

Section: Remodeling Of the Nucleus Through Tasmentioning

confidence: 99%

The significance of telomeric aggregates in the interphase nuclei of tumor cells

Mai

Garini

2006

J of Cellular Biochemistry

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“…Using specific antibodies or nucleic acid probes, several authors have demonstrated the nonrandom distribution and the cell cycle dependent rearrangement of centromeres in interphase nuclei (1,9,13, 36,38). The proximity of specific chromosome centromeres and nucleoli has also been reported (3,14,16,23,24).…”

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Interactive computer‐assisted analysis of chromosome 1 colocalization with nucleoli

et al. 1994

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The applications of DNA cloning and fluorescent in situ hybridization (FISH) techniques have strengthened the hypothesis of an ordered chromatin structure in interphase nuclei, strongly suspected to vary with functional state. The nonrandom distribution of the centromeres and their dynamic rearrangement during the cell cycle have been well documented. A close proximity of specific centromeres to nucleoli has also been reported, but the functional meaning of this association is still unknown. In order to investigate whether the chromosome 1 centromere region to nucleolus association depends on the cell cycle and chromosome status, we combined FISH of probes specific for the lq12 region with Ki-67 nucleolar antigen fluorescent immunocytochemical (FICC) detection on the MCF-7 human breast cancer cell line and on the MRC-5 normal fibroblastic cell line. Both FISH and FICC signals were interactively localized in a one-step fluorescent microscopic observation and further analyzed using the Highly Optimized Microscope Environment (HOME) graphics microscope workstation, which provided computerized interactive marking of lq12 to nucleolus associations (lql2-nu) at the individual nucleus and nucleolus levels. This study confirms that centromeric regions, other than those adjacent to the major ribosomal cistrons, contribute to the perinucleolar chromatin and demonstrate that, during the cell cycle, the heterochromatic band lq12 is dynamically rearranged with regard to both the nuclear volume and the nucleoli. A relationship between the association of the chromosome 1 pericentromeric region with nucleoli and the nucleolar transcriptional activity is also strongly suggested. Key terms: Combination of FISH and immunocytochemistry, functional genome mapping, chromosome 1, nucleolus, interactive cytometry, HOME systemThe recent applications of DNA cloning and fluorescent in situ hybridization (FISH) techniques have strengthened the hypothesis of an ordered chromatin structure, strongly suspected to vary with functional state, in the interphase nuclei (2,(20)(21)(22)(23)27,29,32). Using specific antibodies or nucleic acid probes, several authors have demonstrated the nonrandom distribution and the cell cycle dependent rearrangement of centromeres in interphase nuclei (1,9,13,36,38). The proximity of specific chromosome centromeres and nucleoli has also been reported (3,14,16,23,24). The short distance between the secondary [nucleolus organizer regions (NORs)] and the primary (centromeres) constrictions in acrocentric chromosomes is most likely responsible for the centromere to nucleolus proximity, which is the case for chromosomes 13,14,15,21, and 22 bearing the ribosomal DNA in humans. However, this hypothesis does not apply when chromosomes such as 1 and 9 are involved (23,331 and when the number of chromosome to nucleolus associations is higher than the number of NOR-bearing chromosomes (25).The aim of this work was thus to investigate whether the chromosome 1 centromeric region to nucleolus association depends on the cel...

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Cell Cycle-Dependent Distribution of Telomeres, Centromeres, and Chromosome-Specific Subsatellite Domains in the Interphase Nucleus of Mouse Lymphocytes

Cited by 154 publications

References 0 publications

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

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

The significance of telomeric aggregates in the interphase nuclei of tumor cells

Interactive computer‐assisted analysis of chromosome 1 colocalization with nucleoli

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