Rapid telomere motions in live human cells analyzed by highly time-resolved microscopy

BackgroundTelomeres cap chromosome ends and protect the genome. We studied individual telomeres in live human cancer cells. In capturing telomere motions using quantitative imaging to acquire complete high-resolution three-dimensional datasets every second for 200 seconds, telomere dynamics were systematically analyzed.ResultsThe motility of individual telomeres within the same cancer cell nucleus was widely heterogeneous. One class of internal heterochromatic regions of chromosomes analyzed moved more uniformly and showed less motion and heterogeneity than telomeres. The single telomere analyses in cancer cells revealed that shorter telomeres showed more motion, and the more rapid telomere motions were energy dependent. Experimentally increasing bulk telomere length dampened telomere motion. In contrast, telomere uncapping, but not a DNA damaging agent, methyl methanesulfonate, significantly increased telomere motion.ConclusionNew methods for seconds-scale, four-dimensional, live cell microscopic imaging and data analysis, allowing systematic tracking of individual telomeres in live cells, have defined a previously undescribed form of telomere behavior in human cells, in which the degree of telomere motion was dependent upon telomere length and functionality.

[1]  R. Peters.  Nucleo‐cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis. , 1984, The EMBO journal.

[2]  John W. Sedat,et al.  Multiple regimes of constrained chromosome motion are regulated in the interphase Drosophila nucleus , 2001, Current Biology.

[3]  Carol W. Greider,et al.  The telomere terminal transferase of tetrahymena is a ribonucleoprotein enzyme with two kinds of primer specificity , 1987, Cell.

[4]  Frank H. Wilhelm,et al.  Cell aging in relation to stress arousal and cardiovascular disease risk factors , 2006, Psychoneuroendocrinology.

[5]  T. de Lange,et al.  Shelterin: the protein complex that shapes and safeguards human telomeres. , 2005, Genes & development.

[6]  E. Blackburn,et al.  Human cancer cells harbor T-stumps, a distinct class of extremely short telomeres. , 2007, Molecular cell.

[7]  S. Yusuf,et al.  Effect of potentially modifiable risk factors associated with myocardial infarction in 52 countries (the INTERHEART study): case-control study , 2004, The Lancet.

[8]  P. Gennes,et al.  Reptation of a Polymer Chain in the Presence of Fixed Obstacles , 1971 .

[9]  Tom Misteli,et al.  Maintenance of Stable Heterochromatin Domains by Dynamic HP1 Binding , 2003, Science.

[10]  D. Agard,et al.  Chromosome elasticity and mitotic polar ejection force measured in living Drosophila embryos by four-dimensional microscopy-based motion analysis , 2001, Current Biology.

[11]  D. Malkin,et al.  Younger age of cancer initiation is associated with shorter telomere length in Li-Fraumeni syndrome. , 2007, Cancer research.

[12]  A. Murray,et al.  Interphase chromosomes undergo constrained diffusional motion in living cells , 1997, Current Biology.

[13]  E. Blackburn,et al.  Cellular and Gene Expression Responses Involved in the Rapid Growth Inhibition of Human Cancer Cells by RNA Interference-mediated Depletion of Telomerase RNA* , 2005, Journal of Biological Chemistry.

[14]  Wendy A Bickmore,et al.  Chromatin Motion Is Constrained by Association with Nuclear Compartments in Human Cells , 2002, Current Biology.

[15]  M. Gustafsson,et al.  Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy , 2008, Science.

[16]  Carol W. Greider,et al.  Identification of a specific telomere terminal transferase activity in tetrahymena extracts , 1985, Cell.

[17]  S. Bacchetti,et al.  Reprogramming of telomerase by expression of mutant telomerase RNA template in human cells leads to altered telomeres that correlate with reduced cell viability , 1997, Molecular and cellular biology.

[18]  R. Reddel,et al.  Identification of candidate alternative lengthening of telomeres genes by methionine restriction and RNA interference , 2007, Oncogene.

[19]  V. Andrés,et al.  Aging, telomeres, and atherosclerosis. , 2005, Cardiovascular research.

[20]  P. Lichter,et al.  Mobility of multi-subunit complexes in the nucleus: accessibility and dynamics of chromatin subcompartments , 2005, Histochemistry and Cell Biology.

[21]  K M Hahn,et al.  Dynamic elastic behavior of alpha-satellite DNA domains visualized in situ in living human cells , 1996, The Journal of cell biology.

[22]  T. Misteli,et al.  High mobility of proteins in the mammalian cell nucleus , 2000, Nature.

[23]  Peter Sperisen,et al.  Telomere Length Homeostasis Is Achieved via a Switch between Telomerase- Extendible and -Nonextendible States , 2004, Cell.

[24]  E. Blackburn,et al.  Telomere states and cell fates , 2000, Nature.

[25]  C. Murphy,et al.  Steady-state dynamics of Cajal body components in the Xenopus germinal vesicle , 2003, The Journal of cell biology.

[26]  Roland Eils,et al.  Visualizing telomere dynamics in living mammalian cells using PNA probes , 2003, The EMBO journal.

[27]  E. Blackburn,et al.  Rapid Inhibition of Cancer Cell Growth Induced by Lentiviral Delivery and Expression of Mutant-Template Telomerase RNA and Anti-telomerase Short-Interfering RNA , 2004, Cancer Research.

[28]  Y. Hiraoka,et al.  Cell cycle behavior of human HP1 subtypes: distinct molecular domains of HP1 are required for their centromeric localization during interphase and metaphase , 2003, Journal of Cell Science.

[29]  Karsten Rippe,et al.  Nuclear body movement is determined by chromatin accessibility and dynamics. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[30]  E. Blackburn,et al.  ATM mediates cytotoxicity of a mutant telomerase RNA in human cancer cells. , 2008, Cancer research.

[31]  P. K. Hepler,et al.  Metabolic inhibitors block anaphase A in vivo , 1986, The Journal of cell biology.

[32]  R. Reddel,et al.  Telomerase-negative immortalized human cells contain a novel type of promyelocytic leukemia (PML) body. , 1999, Cancer research.

[33]  E. Blackburn,et al.  Human Rif1 protein binds aberrant telomeres and aligns along anaphase midzone microtubules , 2004, The Journal of cell biology.

[34]  H. Tanke,et al.  Heterogeneity in telomere length of human chromosomes. , 1996, Human molecular genetics.

[35]  Jason R. Swedlow,et al.  Cajal Body dynamics and association with chromatin are ATP-dependent , 2002, Nature Cell Biology.

[36]  P. Lichter,et al.  Histone acetylation increases chromatin accessibility , 2005, Journal of Cell Science.

[37]  Florence Hediger,et al.  Methods for visualizing chromatin dynamics in living yeast. , 2004, Methods in enzymology.

[38]  F. Gage,et al.  In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector , 1996, Science.

[39]  Jason R. Swedlow,et al.  In Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells , 2000, The Journal of cell biology.

[40]  Jean-Christophe Olivo-Marin,et al.  SAGA interacting factors confine sub-diffusion of transcribed genes to the nuclear envelope , 2006, Nature.

[41]  Wendy S. Wright,et al.  (www.interscience.wiley.com) DOI: 10.1002/path.2090 Review Article Hallmarks of telomeres in ageing research , 2022 .

[42]  P. Gennes.  Reptation of a Polymer Chain in the Presence of Fixed Obstacles , 1971 .

[43]  S. Gasser,et al.  Visualizing Chromatin Dynamics in Interphase Nuclei , 2002, Science.

[44]  A. Smogorzewska,et al.  Senescence Induced by Altered Telomere State, Not Telomere Loss , 2002, Science.

[45]  Wen-Lin Kuo,et al.  A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. , 2006, Cancer cell.

[46]  E. Blackburn,et al.  A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis , 1989, Nature.

[47]  Y. Hiraoka,et al.  Activation of the pheromone-responsive MAP kinase drives haploid cells to undergo ectopic meiosis with normal telomere clustering and sister chromatid segregation in fission yeast , 2004, Journal of Cell Science.

[48]  T. Cremer,et al.  Dynamic genome architecture in the nuclear space: regulation of gene expression in three dimensions , 2007, Nature Reviews Genetics.

[49]  R. Eils,et al.  Metabolic-energy-dependent movement of PML bodies within the mammalian cell nucleus , 2002, Nature Cell Biology.

[50]  Susan M. Gasser,et al.  Live Imaging of Telomeres yKu and Sir Proteins Define Redundant Telomere-Anchoring Pathways in Yeast , 2002, Current Biology.