Visualizing telomere dynamics in living mammalian cells using PNA probes
Open Access
- 15 December 2003
- journal article
- research article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 22 (24), 6631-6641
- https://doi.org/10.1093/emboj/cdg633
Abstract
Chromosome ends are protected from degradation by the presence of the highly repetitive hexanucleotide sequence of TTAGGG and associated proteins. These so‐called telomeric complexes are suggested to play an important role in establishing a functional nuclear chromatin organization. Using peptide nucleic acid (PNA) probes, we studied the dynamic behavior of telomeric DNA repeats in living human osteosarcoma U2OS cells. A fluorescent cy3‐labeled PNA probe was introduced in living cells by glass bead loading and was shown to specifically associate with telomeric DNA shortly afterwards. Telomere dynamics were imaged for several hours using digital fluorescence microscopy. While the majority of telomeres revealed constrained diffusive movement, individual telomeres in a human cell nucleus showed significant directional movements. Also, a subfraction of telomeres were shown to associate and dissociate, suggesting that in vivo telomere clusters are not stable but dynamic structures. Furthermore, telomeres were shown to associate with promyelocytic leukemia (PML) bodies in a dynamic manner.Keywords
This publication has 59 references indexed in Scilit:
- Live Imaging of Telomeres: yKu and Sir Proteins Define Redundant Telomere-Anchoring Pathways in YeastCurrent Biology, 2002
- Alternative lengthening of telomeres in mammalian cellsOncogene, 2002
- Alternative lengthening of telomeres is associated with chromosomal instability in osteosarcomasOncogene, 2001
- Telomere Position Effect in Human CellsScience, 2001
- Mutational Analysis of Fibrillarin and Its Mobility in Living Human CellsThe Journal of cell biology, 2000
- Developing a Virtual Reality User Interface (VRUI) for Geographic Information Retrieval on the InternetTransactions in GIS, 1999
- Structure, subnuclear distribution, and nuclear matrix association of the mammalian telomeric complex.The Journal of cell biology, 1996
- Dynamic elastic behavior of alpha-satellite DNA domains visualized in situ in living human cells.The Journal of cell biology, 1996
- Heterogeneity in telomere length of human chromosomesHuman Molecular Genetics, 1996
- The detection of subtelomeric chromosomal rearrangements in idiopathic mental retardationNature Genetics, 1995