Advertisement

Cytotechnology

, Volume 68, Issue 4, pp 1301–1314 | Cite as

Impaired telomerase activity hinders proliferation and in vitro transformation of Penaeus monodon lymphoid cells

  • P. Jayesh
  • S. Vrinda
  • P. Priyaja
  • Rosamma Philip
  • I. S. Bright SinghEmail author
Original Research

Abstract

Retaining terminal transferase activity of telomerase, the ribonucleoprotein enzyme which add telomeric repeats on chromosome end is thought to be required to prevent cellular ageing. Additionally, telomerase considered as a marker for cell proliferation and immortalization in eukaryotes. We examined telomerase activity in tissues and lymphoid cell culture of Penaeus monodon. Along with telomerase activity, telomere repeats and an attempt on identification of telomerase reverse transcriptase (PmTERT) were made. Telomeric repeat amplification protocol revealed that telomerase-dependent telomeric lengthening has been taking place in P. monodon and the adult tissues were retaining this capacity throughout their lifespan with the highest activity in ovary, testis and lymphoid organ. However, telomerase activity could not be detected in lymphoid cells in culture. The canonical telomeric repeats added by telomerase of lymphoid tissue extract were identified as TTAGG, but pentameric repeats GGTTA and AGGTT were also added by the telomerase. PmTERT protein sequence (partial) shared 100 % identity with the TERT sequence of Daphnia pulex, 27 % sequence identity with Purple sea urchin and 24–25 % with Zebra fish. Undetectable telomerase activity in lymphoid cell culture supports the hypothesis that the inadequate telomerase activity or gene expression may be a reason that prevents neoplastic transformation and spontaneous immortalization of the cells in vitro. Thus, it is envisaged that telomerase activation in lymphoid cells may surmount cellular ageing for in vitro transformation and cell line establishment.

Keywords

Telomerase Telomere PmTERT gene Telomerase reverse transcriptase Penaeus monodon Lymphoid cell culture 

Notes

Acknowledgments

This research was supported by Department of Biotechnology, Government of India (BT/PR8050/AAQ/03/289/2006 and BT/PR5126/AAQ/3/591/2012). The first author thanks DBT for Fellowship.

References

  1. Allsopp RC, Vaziri H, Pattersont C, Goldsteint S, Younglai EV, Futcher AB, Greidert CW, Harley CB (1992) Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci USA 89:10114–10118CrossRefGoogle Scholar
  2. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402CrossRefGoogle Scholar
  3. Anchelin M, Murcia L, Alcaraz-Pérez F, García-Navarro EM, Cayuela ML (2011) Behaviour of telomere and telomerase during aging and regeneration in zebrafish. PLoS One 6:e16955CrossRefGoogle Scholar
  4. Armstrong L, Lako M, Lincoln J, Cairns PM, Hole N (2000) mTert expression correlates with telomerase activity during the differentiation of murine embryonic stem cells. Mech Dev 97:109–116CrossRefGoogle Scholar
  5. Balasubramanian S, Kim KH, Ahmad N, Mukhtar H (1999) Activation of telomerase and its association with G1-phase of the cell cycle during UVB-induced skin tumorigenesis in SKH-1 hairless mouse. Oncogene 18:1297–1302CrossRefGoogle Scholar
  6. Belair CD, Yeager TR, Lopez PM, Reznikoff CR (1997) Telomerase activity: a biomarker of cell proliferation, not malignant transformation. Proc Natl Acad Sci USA 94:13677–13682CrossRefGoogle Scholar
  7. Bibby MC (2002) Introduction to telomeres and telomerase. In: Double JA, Thompson MJ (eds) Telomeres and telomerase: methods and protocols. Humana Press, TotowaGoogle Scholar
  8. Blackburn EH (1991) Structure and function of telomeres. Nature 350:569–573CrossRefGoogle Scholar
  9. Blackburn EH (1994) Telomeres: no end in sight. Cell 77:621–623CrossRefGoogle Scholar
  10. Blackburn EH (2001) Switching and signaling at the telomere. Cell 106:661–673CrossRefGoogle Scholar
  11. Bodnar AG, Ouellette M, Frolkis M, Holt ShE, Chiu C-P, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279:349–352CrossRefGoogle Scholar
  12. Bryan TM, Cech TR (1999) Telomerase and the maintenance of chromosome ends. Curr Opin Cell Biol 11:318–324CrossRefGoogle Scholar
  13. Bryan TM, Englezou A, Gupta J, Bacchetti S, Reddell RR (1995) Telomere elongation in immortal human cells without detectable telomerase activity. EMBO J 14:4240–4248Google Scholar
  14. Bryan TM, Englezou A, Dalla-Pozza L, Dunham MA, Reddel RR (1997) Evidence for an alternative mechanism for maintaining telomere length in human tumors and tumor-derived cell lines. Nat Med 3:1271–1274CrossRefGoogle Scholar
  15. Bryan TM, Sperger JM, Chapman KB, Cech TR (1998) Telomerase reverse transcriptase genes identified in Tetrahymena thermophila and Oxytricha trifallax. Proc Natl Acad Sci USA 95:8479–8484CrossRefGoogle Scholar
  16. Cerni C (2000) Telomeres, telomerase, and myc: an update. Mutat Res 462:31–47CrossRefGoogle Scholar
  17. Colbourne JK, Pfrender ME, Gilbert D, Thomas WK, Tucker A, Oakley TH, Tokishita S, Aerts A, Arnold GJ, Basu MK, Bauer DJ, Cáceres CE, Carmel L, Casola C, Choi JH, Detter JC, Dong Q, Dusheyko S, Eads BD, Fröhlich T, Geiler-Samerotte KA, Gerlach D, Hatcher P, Jogdeo S, Krijgsveld J, Kriventseva EV, Kültz D, Laforsch C, Lindquist E, Lopez J, Manak JR, Muller J, Pangilinan J, Patwardhan RP, Pitluck S, Pritham EJ, Rechtsteiner A, Rho M, Rogozin IB, Sakarya O, Salamov A, Schaack S, Shapiro H, Shiga Y, Skalitzky C, Smith Z, Souvorov A, Sung W, Tang Z, Tsuchiya D, Tu H, Vos H, Wang M, Wolf YI, Yamagata H, Yamada T, Ye Y, Shaw JR, Andrews J, Crease TJ, Tang H, Lucas SM, Robertson HM, Bork P, Koonin EV, Zdobnov EM, Grigoriev IV, Lynch M, Boore JL (2011) The ecoresponsive genome of Daphnia pulex. Science 331:555–561Google Scholar
  18. Counter CM, Avilion AA, LeFeuvrel CE, Stewart NG, Greider CW, Harley CB, Bacchettil S (1992) Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:1921–1929Google Scholar
  19. Counter CM, Meyerson M, Eaton EN, Weinberg RA (1997) The catalytic subunit of yeast telomerase. Proc Natl Acad Sci USA 94:9202–9207CrossRefGoogle Scholar
  20. Counter CM, Meyerson M, Eaton EN, Ellisen Leif W, Caddle SD, Haber DA, Weinberg RA (1998) Telomerase activity is restored in human cells by ectopic expression of hTERT (hEST2), the catalytic subunit of telomerase. Oncogene 16:1217–1222CrossRefGoogle Scholar
  21. Delany ME, Daniels LM (2004) The chicken telomerase reverse transcriptase (chTERT): molecular and cytogenetic characterization with a comparative analysis. Gene 339:61–69CrossRefGoogle Scholar
  22. Fitzgerald MS, McKnight TD, Shippen DE (1996) Characterization and developmental patterns of telomerase expression in plants. Proc Natl Acad Sci USA 93:14422–14427CrossRefGoogle Scholar
  23. Forsyth NR, Wright WE, Shay JW (2002) Telomerase and differentiation inmulticellular organisms: turn it off, turn it on, turn it off again. Differentiation 69:188–197CrossRefGoogle Scholar
  24. Frydrychová R, Grossmann P, Trubac P, Vítková M, Marec F (2004) Phylogenetic distribution of TTAGG telomeric repeats in insects. Genome 47:163–178CrossRefGoogle Scholar
  25. Gallardo-Escarate C, Alvarez-Borrego J, Del Rio-Portilla MA, Von Brand-Skopnik E, Cross I, Merlo A, Rebordinos L (2005) Karyotype analysis and chromosomal localization by FISH of ribosomal DNA, telomeric (TTAGGG)n and (GATA)n repeats in Haliotis fulgens and H. corrugata (Archeogastropoda: Haliotidae). J Shellfish Res 24:1153–1159CrossRefGoogle Scholar
  26. Gillis AJ, Schuller AP, Skordalakes E (2008) Structure of the Tribolium castaneum telomerase catalytic subunit TERT. Nature 455:633–638CrossRefGoogle Scholar
  27. Gomes NMV, Shay JW, Wright WE (2010) Telomere biology in Metazoa. FEBS Lett 584:3741–3751CrossRefGoogle Scholar
  28. Gomez D, Mergny JL, Riou JF (2002) Detection of telomerase inhibitors based on G-quadruplex ligands by a modified telomeric repeat amplification protocol assay. Cancer Res 62:3365–3368Google Scholar
  29. Greenberg RA, Allsopp RC, Chin L, Morin GB, DePinho RA (1998) Expression of mouse telomerase reverse transcriptase during development, differentiation and proliferation. Oncogene 16:1723–1730CrossRefGoogle Scholar
  30. Guo W, Okamoto M, Park NH, Lee YM (2001) Cloning and expression of hamster telomerase catalytic subunit cDNA. Int J Mol Med 8:73–78Google Scholar
  31. Harley CB (1991) Telomere loss: mitotic clock or genetic time bomb? Mutat Res 256:271–282CrossRefGoogle Scholar
  32. Harley CB, Futcher AB, Greider CW (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460CrossRefGoogle Scholar
  33. Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636CrossRefGoogle Scholar
  34. Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Cancer Res 25:585–621Google Scholar
  35. He YL, Wu YH, He XN, Liu FJ, He XY, Zhang Y (2009) An immortalized goat mammary epithelial cell line induced with human telomerase reverse transcriptase (hTERT) gene transfer. Theriogenology 71:1417–1424CrossRefGoogle Scholar
  36. Henson JD, Neumann AA, Yeager TR, Reddel RR (2002) Alternative lengthening of telomeres in mammalian cells. Oncogene 21:598–610CrossRefGoogle Scholar
  37. Jayesh P (2013) Development of lymphoid cell culture system from Penaeus monodon and molecular approaches for its transformation. Ph.D. Thesis. Cochin University of Science and Technology, IndiaGoogle Scholar
  38. Jayesh P, Seena J, Singh ISB (2012) Establishment of shrimp cell lines: perception and orientation. Ind J Virol 23:244–251CrossRefGoogle Scholar
  39. Jayesh P, Seena J, Philip R, Singh ISB (2013) A novel medium for the development of in vitro cell culture system from Penaeus monodon. Cytotechnology 65:307–322CrossRefGoogle Scholar
  40. Jayesh P, Philip R, Singh ISB (2015a) Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro. Cytotechnology 67:559–571CrossRefGoogle Scholar
  41. Jayesh P, Philip R, Singh ISB (2015b) Transgene expression in Penaeus monodon cells: evaluation of recombinant baculoviral vectors with shrimp specific hybrid promoters. Cytotechnology. doi: 10.1007/s10616-015-9872-y Google Scholar
  42. Jha AN, Dominquez I, Balajee AS, Hutchinson TH, Dixon DR, Natarajan AT (1995) Localization of a vertebrate telomeric sequence in the chromosomes of two marine worms (phylum Annelida: class Polychaeta). Chromosome Res 3:507–508CrossRefGoogle Scholar
  43. Joffe BI, Solovei IV, Macgregor HC (1996) Ends of chromosomes in Polycelis tenuis (Platyhelminthes) have telomere repeat TTAGGG. Chromosome Res 4:323–324CrossRefGoogle Scholar
  44. Jose S, Jayesh P, Mohandas A, Philip R, Singh ISB (2011) Application of primary haemocyte culture of Penaeus monodon in the assessment of cytotoxicity and genotoxicity of heavy metals and pesticides. Mar Environ Res 71:169–177CrossRefGoogle Scholar
  45. Jose S, Jayesh P, Sudheer NS, Poulose G, Mohandas A, Philip R, Bright ISB (2012) Lymphoid organ cell culture system from Penaeus monodon (Fabricius) as a platform for white spot syndrome virus and shrimp immune-related gene expression. J Fish Dis 35:321–334CrossRefGoogle Scholar
  46. Kim NW, Wu F (1997) Advances in quantification and characterization of telomerase activity by the telomeric repeat amplification protocol (TRAP). Nucleic Acids Res 25:2595–2597CrossRefGoogle Scholar
  47. Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PLC, Coviello GM, Wright WE, Weinrich SL, Shay JW (1994) Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015CrossRefGoogle Scholar
  48. Klapper W, Kuëhne K, Singh KK, Heidorn K, Parwaresch R, Krupp G (1998a) Longevity of lobsters is linked to ubiquitous telomerase expression. FEBS Lett 439:143–146CrossRefGoogle Scholar
  49. Klapper W, Heidorn W, Kuëhne K, Parwaresch R, Krupp G (1998b) Telomerase activity in ‘immortal fish’. FEBS Lett 434:409–412CrossRefGoogle Scholar
  50. Koziol C, Borojevic R, Steffen R, Müller WEG (1998) Sponges (Porifera) model systems to study the shift from immortal to senescent somatic cells: the telomerase activity in somatic cells. Mech Ageing Devt 100:107–120CrossRefGoogle Scholar
  51. Kuramoto M, Ohsumi K, Kishimoto T, Ishikawa F (2001) Identification and analyses of the Xenopus TERT gene that encodes the catalytic subunit of telomerase. Gene 277:101–110CrossRefGoogle Scholar
  52. Lang GH, Wang Y, Nomura N, Matsumura M (2004) Detection of telomerase activity in tissues and primary cultured lymphoid cells of Penaeus japonicus. Mar Biotechnol 6:347–354CrossRefGoogle Scholar
  53. Li Y, Yates JA, Chen JJ-L (2007) Identification and characterization of sea squirt telomerase reverse transcriptase. Gene 400:16–24CrossRefGoogle Scholar
  54. Lundblad V, Blackburn EH (1993) An alternative pathway for yeast telomere maintenance rescues est1 senescence. Cell 73:347–360CrossRefGoogle Scholar
  55. Maxam AM, Gilbert W (1980) Sequencing end-labeled dNA with base-specific chemical cleavages. Methods Enzymol 65:499–560CrossRefGoogle Scholar
  56. Meyerson M, Counter CM, Eaton EN, Ellisen LW, Steiner P, Caddle SD, Ziaugra L, Beijersbergen RL, Davidoff MJ, Liu Q, Bacchetti S, Haber DA, Weinberg RA (1997) hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 90:785–795Google Scholar
  57. Meyne J, Ratliff RL, Moyzis RK (1989) Conservation of the human telomere sequence (TTAGGG)n among vertebrates. Proc Natl Acad Sci USA 86:7049–7053CrossRefGoogle Scholar
  58. Mohan KN, Rani BS, Kulashreshta PS, Kadandale JS (2011) Characterization of TTAGG telomeric repeats, their interstitial occurrence and constitutively active telomerase in the mealybug Planococcus lilacinus (Homoptera; Coccoidea). Chromosoma 120:165–175CrossRefGoogle Scholar
  59. Monti V, Giusti M, Bizzaro D, Manicardi GC, Mandrioli M (2011) Presence of a functional (TTAGG)n telomere-telomerase system in aphids. Chromosome Res 19:625–633CrossRefGoogle Scholar
  60. Morales CP, Holt SE, Ouellette M, Kaur KJ, Yan Y, Wilson KS, White MA, Wright WE, Shay JW (1999) Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nat Genet 21:115–118CrossRefGoogle Scholar
  61. Morin GB (1997) The implications of telomerase biochemistry for human disease. Eur J Cancer 33:750–760CrossRefGoogle Scholar
  62. Muller F, Wicky C, Spicher A, Tobler H (1991) New telomere formation after developmentally regulated chromosomal breakage during the process of chromatin diminution in Ascaris lumbricoides. Cell 67:815–822CrossRefGoogle Scholar
  63. Nakamura TM, Morin GB, Chapman KB, Weinrich SL, Andrews WH, Lingner J, Harley CB, Cech TR (1997) Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955–959CrossRefGoogle Scholar
  64. Nasir L, Gault E, Campbell S, Veeramalai M, Gilbert D, McFarlane R, Munro A, Argyle DJ (2004) Isolation and expression of the reverse transcriptase component of the Canis familiaris telomerase ribonucleoprotein (dogTERT). Gene 336:105–113CrossRefGoogle Scholar
  65. Okazaki S, Tsuchida K, Maekawa H, Ishikawa H, Fujiwara H (1993) Identification of a pentanucleotide telomeric sequence, (TTAGG)n, in the silkworm Bombyx mori and in other insects. Mol Cell Biol 13:1424–1432CrossRefGoogle Scholar
  66. Olovnikov AM (1973) A theory of marginotomy: the incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon. J Theor Biol 41:181–190CrossRefGoogle Scholar
  67. Osanai M, Kojima KK, Futahashi R, Yaguchi S, Fujiwara H (2006) Identification and characterization of the telomerase reverse transcriptase of Bombyx mori (silkworm) and Tribolium castaneum (flour beetle). Gene 376:281–289CrossRefGoogle Scholar
  68. Owen R, Sarkis S, Bodnar A (2007) Developmental pattern of telomerase expression in the sand scallop, Euvola ziczac. Invertebr Biol 126:40–45CrossRefGoogle Scholar
  69. Pardue ML, Danilevskaya ON, Lowenhaupt K, Slot F, Traverse KL (1996) Drosophila telomeres: new views on chromosome evolution. Trends Genet 12:48–52CrossRefGoogle Scholar
  70. Perrem K, Colgin LM, Neumann AA, Yeager TR, Reddel RR (2001) Coexistence of alternative lengthening of telomeres and telomerase in hTERT-transfected GM847 cells. Mol Cell Biol 21:3862–3875CrossRefGoogle Scholar
  71. Piao CQ, Liu L, Zhao YL, Balajee AS, Suzuki M, Hei TK (2005) Immortalization of human small airway epithelial cells by ectopic expression of telomerase. Carcinogenesis 26:725–731CrossRefGoogle Scholar
  72. Podlevsky JD, Bley CJ, Omana RV, Qi X, Chen J (2007) The telomerase database. Nucleic Acids Res 36:339–343CrossRefGoogle Scholar
  73. Rao F, Wang T, Li M, Li Z, Hong N, Zhao H, Yan Y, Lu W, Chen T, Wang W, Lim M, Yuan Y, Liu L, Zeng L, Wei Q, Guan G, Li C, Hong Y (2011) Medaka tert produces multiple variants with differential expression during differentiation in vitro and in vivo. Int J Biol Sci 7:426–439CrossRefGoogle Scholar
  74. Sahara K, Marec F, Traut W (1999) TTAGG telomeric repeats in chromosomes of some insects and other arthropods. Chromosome Res 7:449–460CrossRefGoogle Scholar
  75. Sakai M, Okumura S, Yamamori K (2005) Telomere analysis of pacific abalone Haliotis discus hannai chromosomes by fluorescence in situ hybridization. J Shellfish Res 24:1149–1151CrossRefGoogle Scholar
  76. Sakai M, Okumura S, Onuma K, Senbokuya H, Yamamori K (2007) Identification of a telomere sequence type in three sponge species (Porifera) by fluorescence in situ hybridization analysis. Fish Sci 73:77–80CrossRefGoogle Scholar
  77. Sasaki T, Fujiwara H (2000) Detection and distribution patterns of telomerase activity in insects. Eur J Biochem 267:3025–3031CrossRefGoogle Scholar
  78. Schumpert C, Nelson J, Kim E, Dudycha JL, Patel RC (2015) Telomerase activity and telomere length in Daphnia. PLoS One 10:e0127196. doi: 10.1371/journal.pone.0127196 CrossRefGoogle Scholar
  79. Sharma D, Issac B, Raghava GP, Ramaswamy R (2004) Spectral Repeat Finder (SRF): identification of repetitive sequences using Fourier transformation. Bioinformatics 20:1405–1412CrossRefGoogle Scholar
  80. Singhapol C, Pal D, Czapiewski R, Porika M, Nelson G, Saretzki GC (2013) Mitochondrial telomerase protects cancer cells from nuclear DNA damage and apoptosis. PLoS One 8:e52989. doi: 10.1371/journal.pone.0052989 CrossRefGoogle Scholar
  81. Sköld NH, Asplund MA, Wood CA, Bishop JDD (2011) Telomerase deficiency in a colonial ascidian after prolonged asexual propagation. J Exp Zool B Mol Dev Evol 316:276–283CrossRefGoogle Scholar
  82. Techangamsuwan S, Kreutzer R, Kreutzer M, Imbschweiler I, Rohn K, Wewetzer K, Baumgärtner W (2009) Transfection of adult canine Schwann cells and olfactory ensheathing cells at early and late passage with human TERT differentially affects growth factor responsiveness and in vitro growth. J Neurosci Methods 176:112–120CrossRefGoogle Scholar
  83. Vaziri H, Benchimol S (1998) Reconstitution of telomerase activity in normal human cells leads to elongation of telomeres and extended replication life span. Curr Biol 8:279–282CrossRefGoogle Scholar
  84. Venin SS, Bernard V, Tremblay JP (2000) Telomerase allows the immortalization of T antigen positive DMD myoblasts: a new source of cells for gene transfer application. Gene Ther 7:619–623CrossRefGoogle Scholar
  85. Vitkova M, Krai J, Traut W, Zrzavy J, Marec F (2005) The evolutionary origin of insect telomeric repeats, (TTAGG)n. Chromosome Res 13:145–156CrossRefGoogle Scholar
  86. Vogt G (2012) Hidden treasures in stem cells of indeterminately growing bilaterian invertebrates. Stem Cell Rev 8:305–317. doi: 10.1007/s12015-011-9303-1 CrossRefGoogle Scholar
  87. Vogt G (2012) Ageing and longevity in the Decapoda (Crustacea): a review. Zool Anz 251:1–25CrossRefGoogle Scholar
  88. Wang Y, Guo X (2001) Chromosomal mapping of the vertebrate telomeric sequence (TTAGGG)n in four bivalve molluscs by fluorescence in situ hybridization. J Shellfish Res 20:1187–1190Google Scholar
  89. Wright WE, Piatyszek MA, Rainey WE, Byrd W, Shay JW (1996) Telomerase activity in human germline and embryonic tissues and cells. Dev Genet 18:173–179CrossRefGoogle Scholar
  90. Zielke S, Bodnar A (2010) Telomeres and telomerase activity in scleractinian corals and Symbiodinium spp. Biol Bull 218:113–121Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • P. Jayesh
    • 1
    • 2
  • S. Vrinda
    • 1
  • P. Priyaja
    • 1
  • Rosamma Philip
    • 2
  • I. S. Bright Singh
    • 1
    Email author
  1. 1.National Centre for Aquatic Animal HealthCochin University of Science and TechnologyKochiIndia
  2. 2.Department of Marine Biology, Microbiology and BiochemistryCochin University of Science and TechnologyKochiIndia

Personalised recommendations