#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Increased amounts and stability of telomeric repeat-containing RNA (TERRA) following DNA damage induced by etoposide


Autoři: Bong-Kyeong Oh aff001;  Yoojung Choi aff002;  Jaeman Bae aff003;  Won Moo Lee aff003;  Jeong-Kyu Hoh aff003;  Joong Sub Choi aff001
Působiště autorů: Institute for the Integration of Medicine and Innovative Technology, Hanyang University College of Medicine, Seoul, Korea aff001;  Department of Translational Medicine, Hanyang University Graduate School of Biomedical Science and Engineering, Seoul, Korea aff002;  Department of Obstetrics and Gynecology, Hanyang University College of Medicine, Seoul, Korea aff003
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0225302

Souhrn

Telomeric repeat-containing RNAs (TERRAs) are long noncoding RNAs transcribed from subtelomeres toward telomeric repeat tracts, which have been implicated in telomere protection and heterochromatin formation. Genotoxic stress leads to upregulation of TERRAs. However, the mechanism of DNA damage-mediated TERRA induction remains elusive. Here, we treated HeLa cells with etoposide, a DNA double-strand break-generating agent, for various times and monitored the levels of TERRAs. Etoposide treatment led to a gradual time-dependent increase in TERRAs. Etoposide-mediated induction was evident in many TERRAs arising from various chromosome loci, including 20q and XpYp. Chromatin immunoprecipitation assays revealed no significant changes in the occupancy of RNA polymerase II at telomeres upon etoposide treatment. Interestingly, TERRAs arising from 20q, XpYp, 10q, and 13q degraded at slower rates in cells treated with etoposide, while degradation rates of TERRAs from many loci tested were nearly identical in both etoposide- and mock-treated cells. Telomere damage occurred from early time points of etoposide treatment, but telomere lengths and abundance of telomeric repeat-binding factor 2 (TRF2) at telomeres remained unchanged. In summary, etoposide treatment led to telomere damage and TERRA accumulation, but telomere lengths and TRF2-mediated telomere integrity were maintained. Etoposide-mediated TERRA accumulation could be attributed partly to RNA stabilization. These findings may provide insight into the post-transcriptional regulation of TERRAs in response to DNA damage.

Klíčová slova:

Genetic loci – Telomeres – Telomere length – DNA damage – Ribosomal RNA – Probe hybridization – HeLa cells – Cell hybridization


Zdroje

1. Artandi SE, DePinho RA. Telomeres and telomerase in cancer. Carcinogenesis. 2010;31: 9–18. doi: 10.1093/carcin/bgp268 19887512

2. Palm W, de Lange T. How shelterin protects mammalian telomeres. Annu Rev Genet. 2008;42: 301–334. doi: 10.1146/annurev.genet.41.110306.130350 18680434

3. van Steensel B, Smogorzewska A, de Lange T. TRF2 protects human telomeres from end-to-end fusions. Cell. 1998;92: 401–413. doi: 10.1016/s0092-8674(00)80932-0 9476899

4. Karlseder J, Broccoli D, Dai Y, Hardy S, de Lange T. p53- and ATM-dependent apoptosis induced by telomeres lacking TRF2. Science. 1999;26: 1321–1325.

5. Denchi EL, de Lange T. Protection of telomeres through independent control of ATM and ATR by TRF2 and POT1. Nature. 2007;448: 1068–1071. doi: 10.1038/nature06065 17687332

6. de Lange T, Shiue L, Myers RM, Cox DR, Naylor SL, Killery AM, et al. Structure and variability of human chromosome ends. Mol Cell Biol. 1990;10: 518–527. doi: 10.1128/mcb.10.2.518 2300052

7. Wellinger RJ, Ethier K, Labrecque P, Zakian VA. Evidence for a new step in telomere maintenance. Cell. 1996;85: 423–433. doi: 10.1016/s0092-8674(00)81120-4 8616897

8. d’Adda di Fagagna F, Reaper PM, Clay-Farrace L, Fiegler H, Carr P, Von Zglinicki T, et al. A DNA damage checkpoint response in telomere-initiated senescence. Nature. 2006;426: 194–198.

9. Azzalin CM, Reichenbach P, Khoriauli L, Giulotto E, Lingner J. Telomeric repeat containing RNA and RNA surveillance factors at mammalian chromosome ends. Science. 2007;318: 798–801. doi: 10.1126/science.1147182 17916692

10. Schoeftner S, Blasco M. Developmentally regulated transcription of mammalian telomeres by DNA-dependent RNA polymerase II. Nat Cell Biol. 2008;10: 228–236. doi: 10.1038/ncb1685 18157120

11. Luke B, Panza A, Redon S, Iglesias N, Li Z, Linger J. The Rat1p 59 to 39 exonuclease degrades telomeric repeat-containing RNA and promotes telomere elongation in Saccharomyces cerevisiae. Mol Cell. 2008;32: 465–477. doi: 10.1016/j.molcel.2008.10.019 19026778

12. Bah A, Wischnewski H, Shchepachev V, Azzalin CM. The telomeric transcriptome of Schizosaccharomyces pombe. Nucleic Acids Res. 2012;40: 2995–3005. doi: 10.1093/nar/gkr1153 22139915

13. Montero JJ, López de Silanes I, Graña O, Blasco MA. Telomeric RNAs are essential to maintain telomeres. Nat Commun. 2016;7: 12534. doi: 10.1038/ncomms12534 27531349

14. Chu HP, Cifuentes-Rojas C, Kesner B, Aeby E, Lee HG, Wei C, et al. TERRA RNA antagonizes ATRX and protects telomeres. Cell. 2017;170: 86–101. doi: 10.1016/j.cell.2017.06.017 28666128

15. Lopez de Silanes I, Grana O, De Bonis ML, Dominguez O, Pisano DG, Blasco MA. Identification of TERRA locus unveils a telomere protection role through association to nearly all chromosomes. Nat Commun. 2014;5: 4723. doi: 10.1038/ncomms5723 25182072

16. Deng Z, Norseen J, Wiedmer A, Riethman H, Lieberman PM. TERRA RNA binding to TRF2 facilitates heterochromatin formation and ORC recruitment at telomeres. Mol Cell. 2009;35: 403–413. doi: 10.1016/j.molcel.2009.06.025 19716786

17. Flynn RL, Centore RC, O’Sullivan RJ, Rai R, Tse A, Songyang Z, et al. TERRA and hnRNPA1 orchestrate an RPA-to-POT1 switch on telomeric single-stranded DNA. Nature. 2011;471: 532–536. doi: 10.1038/nature09772 21399625

18. Flynn RL, Cox KE, Jeitany M, Wakimoto H, Bryll AR, Ganem NJ, et al. Alternative lengthening of telomeres renders cancer cells hypersensitive to ATR inhibitors. Science. 2015;347: 273–277. doi: 10.1126/science.1257216 25593184

19. Porro A, Feuerhahn S, Delafontaine J, Riethman H, Rougemont J, Lingner J. Functional characterization of the TERRA transcriptome at damaged telomeres. Nat Commun. 2014;5: 5379–5391. doi: 10.1038/ncomms6379 25359189

20. Deng Z, Wang Z, Stong N, Plasschaert R, Moczan A, Chen HS, et al. A role for CTCF and cohesin in subtelomere chromatin organization, TERRA transcription, and telomere end protection. EMBO J. 2012;31: 4165–4178. doi: 10.1038/emboj.2012.266 23010778

21. Feretzaki M, Renck Nunes P, Lingner J. Expression and differential regulation of human TERRA at several chromosome ends. RNA. 2019.

22. Koskas S, Decottignies A, Dufour S, Pezet M, Verdel A, Vourc’h C, et al. Heat shock factor 1 promotes TERRA transcription and telomere protection upon heat stress. Nucleic Acids Res. 2017;45: 6321–6333. doi: 10.1093/nar/gkx208 28369628

23. Baker AM, Fu Q, Hayward W, Victoria S, Pedroso IM, Stuart M, et al. The telomere binding protein TRF2 induces chromatin compaction. PLoS ONE. 2011;6: e19124. doi: 10.1371/journal.pone.0019124 21526145

24. Poulet A, Pisano S, Faivre-Moskalenko C, Pei B, Tauran Y, Haftek-Terreau Z, et al. The N-terminal domains of TRF1 and TRF2 regulate their ability to condense telomeric DNA. Nucleic Acids Res. 2012;40: 2566–2576. doi: 10.1093/nar/gkr1116 22139926

25. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory; 1989.

26. Oh BK, Chae KJ, Park C, Kim K, Lee WJ, Han KH, et al. Telomere shortening and telomerase reactivation in dysplastic nodules of human hepatocarcinogenesis. J Hepatol. 2003;39: 786–792. doi: 10.1016/s0168-8278(03)00395-7 14568262

27. Kruk PA, Rampino NJ, Bohr VA. DNA damage and repair in telomeres: relation to aging. PNAS. 1995;92: 258–262. doi: 10.1073/pnas.92.1.258 7816828

28. Feretzaki M, Lingner J. A practical qPCR approach to detect TERRA, the elusive telomeric repeat-containing RNA. Methods. 2017;114: 39–45. doi: 10.1016/j.ymeth.2016.08.004 27530378

29. Brown T, Mackey K, Du T. Analysis of RNA by Northern and slot blot hybridization. Curr Protoc Mol Biol. 2004;67: 4.9.1–4.9.19.

30. Sun BK, Deaton AM, Lee JT. A transient heterochromatic state in Xist preempts X-inactivation choice without RNA stabilization. Mol Cell. 2006;21: 617–628. doi: 10.1016/j.molcel.2006.01.028 16507360

31. Porro A, Feuerhahn S, Lingner J. TERRA-reinforced association of LSD1 with MRE11 promotes processing of uncapped telomeres. Cell Rep. 2014;6: 765–776. doi: 10.1016/j.celrep.2014.01.022 24529708

32. Montecucco A, Biamonti G. Cellular response to etoposide treatment. Cancer Lett. 2007;252: 9–18. doi: 10.1016/j.canlet.2006.11.005 17166655

33. Klapper W, Qian W, Schulte C, Parwaresch R. DNA damage transiently increases TRF2 mRNA expression and telomerase activity. Leukemia. 2003;17: 2007–2015. doi: 10.1038/sj.leu.2403086 14513051

34. Liu M, Hales BF, Robaire B. Effects of four chemotherapeutic agents, bleomycin, etoposide, cisplatin, and cyclophosphamide, on DNA damage and telomeres in a mouse spermatogonial cell line. Biol Reprod. 2014;90: 1–10.

35. el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al. WAF1, a potential mediator of p53 tumor suppression. Cell. 1993;75: 817–825. doi: 10.1016/0092-8674(93)90500-p 8242752

36. Gu Y, Turck CW, Morgan DO. Inhibition of CDK2 activity in vivo by an associated 20K regulatory subunit. Nature. 1993;366: 707–710. doi: 10.1038/366707a0 8259216

37. Burden DA, Kingma PS, Froelich-Ammon SJ, Bjornsti MA, Patchan MW, Thompson RB, et al. Topoisomerase II. Etoposide interactions direct the formation of drug-induced enzyme-DNA cleavage complexes. J Biol Chem. 1996;271: 29238–29244. doi: 10.1074/jbc.271.46.29238 8910583

38. Porro A, Feuerhahn S, Reichenbach P, Lingner J. Molecular dissection of telomeric repeat-containing RNA biogenesis unveils the presence of distinct and multiple regulatory pathways. Mol Cell Biol. 2010;20: 4808–4817.


Článok vyšiel v časopise

PLOS One


2019 Číslo 11
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#