Abstract
We describe a method that assesses the impact of specific mutations of TP53 and genomic instability on gene expression of the most important genes involved in telomere length and structure homeostasis. The approaches consist of using a reverse transcriptase method and a quantitative PCR that were applied to isogenic cell lines from a colon cancer.
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References
Benetti R, Garcia-Cao M, Blasco MA (2007) Telomere length regulates the epigenetic status of mammalian telomeres and subtelomeres. Nat Genet 39:243–250. https://doi.org/10.1038/ng1952
De Lange T, Shiue L, Myers RM et al (1990) Structure and variability of human chromosome ends. Mol Cell Biol 10:518–527
Moyzis RK, Buckingham JM, Cram LS et al (1988) A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc Natl Acad Sci U S A 85:6622–6626
Chong L, van Steensel B, Broccoli D et al (1995) A human telomeric protein. Science 270:1663–1667
Lejnine S, Makarov VL, Langmore JP (1995) Conserved nucleoprotein structure at the ends of vertebrate and invertebrate chromosomes. Proc Natl Acad Sci U S A 92:2393–2397
Counter CM, Avilion AA, LeFeuvre CE et al (1992) Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:1921–1929
Nakamura TM, Morin GB, Chapman KB et al (1997) Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955
Zhao YM, Li JY, Lan JP et al (2008) Cell cycle dependent telomere regulation by telomerase in human bone marrow mesenchymal stem cells. Biochem Biophys Res Commun 369:1114–1119. https://doi.org/10.1016/j.bbrc.2008.03.011
Stansel RM, de Lange T, Griffith JD (2001) T-loop assembly in vitro involves binding of TRF2 near the 3′ telomeric overhang. EMBO J 20:5532
Munoz-Jordan JL, Cross GA, de Lange T, Griffith JD (2001) T-loops at trypanosome telomeres. EMBO J 20:579
De Lange T (2004) T-loops and the origin of telomeres. Nat Rev Mol Cell Biol 5:323
Sfeir A, de Lange T (2012) Removal of shelterin reveals the telomere end-protection problem. Science 336:593–597. https://doi.org/10.1126/science.1218498;
Celli GB, Denchi EL, de Lange T (2006) Ku70 stimulates fusion of dysfunctional telomeres yet protects chromosome ends from homologous recombination. Nat Cell Biol 8:885–890. https://doi.org/10.1038/ncb1444
Denchi EL, de Lange T (2007) Protection of telomeres through independent control of ATM and ATR by TRF2 and POT1. Nature 448:1068–1071. https://doi.org/10.1038/nature06065
Sfeir A, Kosiyatrakul ST, Hockemeyer D et al (2009) Mammalian telomeres resemble fragile sites and require TRF1 for efficient replication. Cell 138:90–103. https://doi.org/10.1016/j.cell.2009.06.021
Takai KK, Kibe T, Donigian JR et al (2011) Telomere protection by TPP1/POT1 requires tethering to TIN2. Mol Cell 44:647–659. https://doi.org/10.1016/j.molcel.2011.08.043;
Maciejowski J, de Lange T (2017) Telomeres in cancer: tumour suppression and genome instability. Nat Rev Mol Cell Biol 18:175–186. https://doi.org/10.1038/nrm.2016.171
Romanov SR, Kozakiewicz BK, Holst CR et al (2001) Normal human mammary epithelial cells spontaneously escape senescence and acquire genomic changes. Nature 409:633–637. https://doi.org/10.1038/35054579
Davoli T, de Lange T (2012) Telomere-driven tetraploidization occurs in human cells undergoing crisis and promotes transformation of mouse cells. Cancer Cell 21:765–776. https://doi.org/10.1016/j.ccr.2012.03.044
Davoli T, Denchi EL, de Lange T (2010) Persistent telomere damage induces bypass of mitosis and tetraploidy. Cell 141:81–93. https://doi.org/10.1016/j.cell.2010.01.031
Yates LR, Campbell PJ (2012) Evolution of the cancer genome. Nat Rev Genet 13:795–806. https://doi.org/10.1038/nrg3317
Righolt C, Mai S (2012) Shattered and stitched chromosomes-chromothripsis and chromoanasynthesis-manifestations of a new chromosome crisis? Genes Chromosomes Cancer 51:975–981. https://doi.org/10.1002/gcc.21981
Rausch T, Jones DTW, Zapatka M et al (2012) Genome sequencing of pediatric medulloblastoma links catastrophic DNA rearrangements with TP53 mutations. Cell 148:59–71. https://doi.org/10.1016/j.cell.2011.12.013
Samassekou O, Bastien N, Lichtensztejn D et al (2014) Different TP53 mutations are associated with specific chromosomal rearrangements, telomere length changes, and remodeling of the nuclear architecture of telomeres. Genes Chromosomes Cancer 53:934–950. https://doi.org/10.1002/gcc.22205
Pocard M, Chevillard S, Villaudy J et al (1996) Different p53 mutations produce distinct effects on the ability of colon carcinoma cells to become blocked at the G1/S boundary after irradiation. Oncogene 12:875–882
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Samassekou, O., Bastien, N., Yan, J., Mai, S., Drouin, R. (2018). Expression of Genes Associated with Telomere Homeostasis in TP53 Mutant LoVo Cell Lines as a Model for Genomic Instability. In: Pellestor, F. (eds) Chromothripsis. Methods in Molecular Biology, vol 1769. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7780-2_16
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DOI: https://doi.org/10.1007/978-1-4939-7780-2_16
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