Kings and queens of termites are endowed with an extraordinary longevity coupled with lifelong fecundity. We recently reported that termite kings and queens display a dramatically increased enzymatic activity and abundance of telomerase in their somatic organs when compared to short-lived workers and soldiers. We hypothesized that this telomerase activation may represent a noncanonical pro-longevity function, independent of its canonical role in telomere maintenance. Here, we explore this avenue and investigate whether the presumed noncanonical role of telomerase may be due to alternative splicing of the catalytic telomerase subunit TERT and whether the subcellular localization of TERT isoforms differs among organs and castes in the termite Prorhinotermes simplex. We empirically confirm the expression of four in silico predicted splice variants (psTERT1-A, psTERT1-B, psTERT2-A, psTERT2-B), defined by N-terminal splicing implicating differential localizations, and C-terminal splicing giving rise to full-length and truncated isoforms. We show that the transcript proportions of the psTERT are caste- and tissue-specific and that the extranuclear full-length isoform TERT1-A is relatively enriched in the soma of neotenic kings and queens compared to their gonads and to the soma of workers. We also show that extranuclear TERT protein quantities are significantly higher in the soma of kings and queens compared to workers, namely due to the cytosolic TERT. Independently, we confirm by microscopy the extranuclear TERT localization in somatic organs. We conclude that the presumed pleiotropic action of telomerase combining the canonical nuclear role in telomere maintenance with extranuclear functions is driven by complex TERT splicing.

Faculty of Science Charles University Prague Czech Republic

Faculty of Science University of South Bohemia České Budějovice Czech Republic

Institute of Entomology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic

Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Prague Czech Republic

See more in PubMed

Ahmed, S. , Passos, J.F. , Birket, M.J. , Beckmann, T. , Brings, S. , Peters, H. et al. (2008) Telomerase does not counteract telomere shortening but protects mitochondrial function under oxidative stress. Journal of Cell Science, 121, 1046–1053. PubMed

Anders, S. , Reyes, A. and Huber, W. (2012) Detecting differential usage of exons from RNA‐seq data. Genome Research, 22, 2008–2017. PubMed PMC

Avin, B.A. , Umbricht, C.B. and Zeiger, M.A. (2016) Human telomerase reverse transcriptase regulation by DNA methylation, transcription factor binding and alternative splicing (Review). International Journal of Oncology, 49, 2199–2205. PubMed PMC

Banik, S.S.R. , Guo, C. , Smith, A.C. , Margolis, S.S. , Richardson, D.A. , Tirado, C.A. et al. (2002) C‐terminal regions of the human telomerase catalytic subunit essential for in vivo enzyme activity. Molecular and Cellular Biology, 22, 6234–6246. PubMed PMC

Bernadou, A. , Kramer, B.H. and Korb, J. (2021) Major evolutionary transitions in social insects, the importance of worker sterility and life history trade‐offs. Frontiers in Ecology and Evolution, 9, 732907.

Blackburn, E.H. (2005) Telomeres and telomerase: their mechanisms of action and the effects of altering their functions. FEBS Letters, 579, 859–862. PubMed

Bolger, A.M. , Lohse, M. and Usadel, B. (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 30, 2114–2120. PubMed PMC

Colgin, L.M. , Wilkinson, C. , Englezou, A. , Kilian, A. , Robinson, M.O. and Reddel, R.R. (2000) The hTERTα splice variant is a dominant negative inhibitor of telomerase activity. Neoplasia, 2, 426–432. PubMed PMC

Cong, Y. and Shay, J.W. (2008) Actions of human telomerase beyond telomeres. Cell Research, 18, 725–732. PubMed

Corona, M. , Hughes, K.A. , Weaver, D.B. and Robinson, G.E. (2005) Gene expression patterns associated with queen honey bee longevity. Mechanisms of Ageing and Development, 126, 1230–1238. PubMed

Corona, M. , Velarde, R.A. , Remolina, S. , Moran‐Lauter, A. , Wang, Y. , Hughes, K.A. et al. (2007) Vitellogenin, juvenile hormone, insulin signaling, and queen honey bee longevity. Proceedings of the National Academy of Sciences USA, 104, 7128–7133. PubMed PMC

Denham, J. (2023) Canonical and extra‐telomeric functions of telomerase: implications for healthy ageing conferred by endurance training. Aging Cell, 22, e13836. PubMed PMC

Ding, D. , Zhou, J. , Wang, M. and Cong, Y.S. (2013) Implications of telomere‐independent activities of telomerase reverse transcriptase in human cancer. FEBS Journal, 280, 3205–3211. PubMed

Dobin, A. , Davis, C.A. , Schlesinger, F. , Drenkow, J. , Zaleski, C. , Jha, S. et al. (2013) STAR: ultrafast universal RNA‐seq aligner. Bioinformatics, 29, 15–21. PubMed PMC

Eitan, E. , Braverman, C. , Tichon, A. , Gitler, D. , Hutchison, E.R. , Mattson, M.P. et al. (2016) Excitotoxic and radiation stress increase TERT levels in the mitochondria and cytosol of cerebellar Purkinje neurons. The Cerebellum, 15, 509–517. PubMed PMC

Elsner, D. , Meusemann, K. and Korb, J. (2018) Longevity and transposon defense, the case of termite reproductives. Proceedings of the National Academy of Sciences USA, 115, 5504. PubMed PMC

Frydrychová, R. , Grossmann, P. , Trubac, P. , Vítková, M. and Marec, F.E. (2004) Phylogenetic distribution of TTAGG telomeric repeats in insects. Genome, 47, 163–178. PubMed

Frydrychová, R.Č. (2023) Telomerase as a possible key to bypass reproductive cost. Molecular Ecology, 32, 2134–2143. PubMed

Fu, W. , Killen, M. , Culmsee, C. , Dhar, S. , Pandita, T.K. and Mattson, M.P. (2000) The catalytic subunit of telomerase is expressed in developing brain neurons and serves a cell survival‐promoting function. Journal of Molecular Neurosciences, 14, 3–15. PubMed

Fu, W. , Lu, C. and Mattson, M.P. (2002) Telomerase mediates the cell survival‐promoting actions of brain‐derived neurotrophic factor and secreted amyloid precursor protein in developing hippocampal neurons. Journal of Neurosciences, 22, 10710–10719. PubMed PMC

Greenberg, A.R. (2005) Telomeres, crisis and cancer. Current Molecular Medicine, 5, 213–218. PubMed

Greider, C.W. and Blackburn, E.H. (1985) Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell, 43, 405–413. PubMed

Haendeler, J. , Drose, S. , Buchner, N. , Jakob, S. , Altschmied, J. , Goy, C. et al. (2009) Mitochondrial telomerase reverse transcriptase binds to and protects mitochondrial DNA and function from damage. Arteriosclerosis, Thrombosis, and Vascular Biology, 29, 929–935. PubMed

Haendeler, J. , Hoffmann, J. , Brandes, R.P. , Zeiher, A.M. and Dimmeler, S. (2003) Hydrogen peroxide triggers nuclear export of telomerase reverse transcriptase via Src kinase family‐dependent phosphorylation of tyrosine 707. Molecular and Cellular Biology, 23, 4598–4610. PubMed PMC

Haroon, Ma, X.M. , Li, Y.X. , Zhang, H.X. , Liu, Q. , Su, X.H. et al. (2020) Transcriptomic evidence that insulin signalling pathway regulates the ageing of subterranean termite castes. Scientific Reports, 10, 8187. PubMed PMC

Heidinger, B.J. , Blount, J.D. , Boner, W. , Griffiths, K. , Metcalfe, N.B. and Monaghan, P. (2012) Telomere length in early life predicts lifespan. Proceedings of the National Academy of Sciences USA, 109, 1743–1748. PubMed PMC

Hellemans, S. and Hanus, R. (2024) Termite primary queen—ancestral, but highly specialized eusocial phenotype. Current Opinion in Insect Science, 61, 101157. PubMed

Hrdličková, R. , Nehyba, J. and Bose, H.R. (2012a) Alternatively spliced telomerase reverse transcriptase variants lacking telomerase activity stimulate cell proliferation. Molecular and Cellular Biology, 32, 4283–4296. PubMed PMC

Hrdličková, R. , Nehyba, J. , Lim, S.L. , Grützner, F. and Bose, H.R. (2012b) Insights into the evolution of mammalian telomerase: Platypus TERT shares similarities with genes of birds and other reptiles and localizes on sex chromosomes. BMC Genomics, 13, 216. PubMed PMC

Huard, S. , Moriarty, T.J. and Autexier, C. (2003) The C terminus of the human telomerase reverse transcriptase is a determinant of enzyme processivity. Nucleic Acids Research, 31, 4059–4070. PubMed PMC

Iannilli, F. , Zalfa, F. , Gartner, A. , Bagni, C. and Dotti, C.G. (2013) Cytoplasmic TERT associates to RNA granules in fully mature neurons: role in the translational control of the cell cycle inhibitor p15INK4B. PLoS ONE, 8, e66602. PubMed PMC

Indran, I.R. , Hande, M.P. and Pervaiz, S. (2011) hTERT overexpression alleviates intracellular ROS production, improves mitochondrial function, and inhibits ROS‐mediated apoptosis in cancer cells. Cancer Research, 71, 266–276. PubMed

Jaskelioff, M. , Muller, F.L. , Paik, J.H. , Thomas, E. , Jiang, S. , Adams, A.C. et al. (2011) Telomerase reactivation reverses tissue degeneration in aged telomerase‐deficient mice. Nature, 469, 102–106. PubMed PMC

Jemielity, S. , Kimura, M. , Parker, K.M. , Parker, J.D. , Cao, X. , Aviv, A. et al. (2007) Short telomeres in short‐lived males: what are the molecular and evolutionary causes? Aging Cell, 6, 225–233. PubMed PMC

Keller, L. (1998) Queen lifespan and colony characteristics in ants and termites. Insectes Sociaux, 45, 235–246.

Keller, L. and Genoud, M. (1997) Extraordinary lifespans in ants: a test of evolutionary theories of ageing. Nature, 389, 958–960.

Klapper, W. , Shin, T. and Mattson, M.P. (2001) Differential regulation of telomerase activity and TERT expression during brain development in mice. Journal of Neuroscience Research, 64, 252–260. PubMed

Korandová, M. and Frydrychová, R.Č. (2016) Activity of telomerase and telomeric length in Apis mellifera . Chromosoma, 125, 405–411. PubMed

Korandová, M. , Krůček, T. , Vrbová, K. and Frydrychová, R.Č. (2014) Distribution of TTAGG‐specific telomerase activity in insects. Chromosome Research, 22, 495–503. PubMed

Korb, J. and Heinze, J. (2021) Ageing and sociality: why, when and how does sociality change ageing patterns? Philosophical Transactions of the Royal Society B—Biological Sciences, 376, 20190727. PubMed PMC

Korb, J. , Meusemann, K. , Aumer, D. , Bernadou, A. , Elsner, D. , Feldmeyer, B. et al. (2021) Comparative transcriptomic analysis of the mechanisms underpinning ageing and fecundity in social insects. Philosophical Transactions of the Royal Society B—Biological Sciences, 376, 20190728. PubMed PMC

Korb, J. and Thorne, B. (2017) Sociality in termites. In Comparative Social Evolution (eds. Rubenstein D.R. & Abbot P.), pp. 124–153. Cambridge University Press, Cambridge.

Koubová, J. and Frydrychová, R.Č. (2021) Telomerase‐positive somatic tissues of honeybee queens (Apis mellifera) display no DNA replication. Cytogenetic and Genome Research, 161, 470–475. PubMed

Koubová, J. , Jehlík, T. , Kodrík, D. , Sábová, M. , Šima, P. , Sehadová, H. et al. (2019) Telomerase activity is upregulated in the fat bodies of pre‐diapause bumblebee queens (Bombus terrestris). Insect Biochemistry and Molecular Biology, 115, 103241. PubMed

Koubová, J. , Pangrácová, M. , Jankásek, M. , Lukšan, O. , Jehlík, T. , Brabcová, J. et al. (2021) Long‐lived termite kings and queens activate telomerase in somatic organs. Proceedings of the Royal Society B—Biological Sciences, 288, 20210511. PubMed PMC

Kramer, B.H. , Nehring, V. , Buttstedt, A. , Heinze, J. , Korb, J. , Libbrecht, R. et al. (2021) Oxidative stress and senescence in social insects: a significant but inconsistent link? Philosophical Transactions of the Royal Society B ‐ Biological Sciences, 376, 20190732. PubMed PMC

Kuznetsova, V. , Grozeva, S. and Gokhman, V. (2020) Telomere structure in insects: a review. Journal of Zoological Systematics and Evolutionary Research, 58, 127–158.

Lai, A.G. , Pouchkina‐Stantcheva, N. , Di Donfrancesco, A. , Kildisiute, G. , Sahu, S. and Aboobaker, A.A. (2017) The protein subunit of telomerase displays patterns of dynamic evolution and conservation across different metazoan taxa. BMC Evolutionary Biology, 17, 107. PubMed PMC

Levine, J.D. , Šauman, I. , Imbalzano, M. , Reppert, S.M. and Jackson, F. (1995) Period protein from the giant silkmoth Antheraea pernyi functions as a circadian clock element in Drosophila melanogaster . Neuron, 15, 147–157. PubMed

Liao, Y. , Smyth, G.K. and Shi, W. (2014) featureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics, 30, 923–930. PubMed

Lin, S. , Elsner, D. , Ams, L. , Korb, J. and Rosengaus, R. (2024) A genetic toolkit underlying the queen phenotype in termites with totipotent workers. Scientific Reports, 14, 2214. PubMed PMC

Lin, S. , Werle, J. and Korb, J. (2021) Transcriptomic analyses of the termite, Cryptotermes secundus, reveal a gene network underlying a long lifespan and high fecundity. Communications Biology, 4, 384. PubMed PMC

Liu, K. , Hodes, R.J. and Weng, N. (2001) Cutting edge: telomerase activation in human T lymphocytes does not require increase in telomerase reverse transcriptase (hTERT) protein but is associated with hTERT phosphorylation and nuclear translocation. Journal of Immunology, 166, 4826–4830. PubMed

Mason, J.M. , Randall, T.A. and Frydrychova, R.C. (2016) Telomerase lost? Chromosoma, 125, 65–73. PubMed PMC

Monroy Kuhn, J.M. and Korb, J. (2016) Editorial overview: social insects: aging and the re‐shaping of the fecundity/longevity trade‐off with sociality. Current Opinion in Insect Science, 16, 7–10. PubMed

Monroy Kuhn, J.M. , Meusemann, K. and Korb, J. (2019) Long live the queen, the king and the commoner? Transcript expression differences between old and young in the termite Cryptotermes secundus . PLoS ONE, 14, e0210371. PubMed PMC

Monroy Kuhn, J.M. , Meusemann, K. and Korb, J. (2021) Disentangling the aging gene expression network of termite queens. BMC Genomics, 22, 339. PubMed PMC

Monti, V. , Giusti, M. , Bizzaro, D. , Manicardi, G.C. and Mandrioli, M. (2011) Presence of a functional (TTAGG) n telomere‐telomerase system in aphids. Chromosome Research, 19, 625–633. PubMed

Parker, J.D. , Parker, K.M. , Sohal, B.H. , Sohal, R.S. and Keller, L. (2004) Decreased expression of Cu‐Zn superoxide dismutase 1 in ants with extreme lifespan. Proceedings of the National Academy of Sciences USA, 101, 3486–3489. PubMed PMC

Penev, A. , Bazley, A. , Shen, M. , Boeke, J.D. , Savage, S.A. and Sfeir, A. (2021) Alternative splicing is a developmental switch for hTERT expression. Molecular Cell, 81, 2349–2360.e6. PubMed PMC

Post, F. , Bornberg‐Bauer, E. , Vasseur‐Cognet, M. and Harrison, M.C. (2023) More effective transposon regulation in fertile, long‐lived termite queens than in sterile workers. Molecular Ecology, 32, 369–380. PubMed

Rau, V. , Flatt, T. and Korb, J. (2023) The remoulding of dietary effects on the fecundity/longevity trade‐off in a social insect. BMC Genomics, 24, 244. PubMed PMC

Roisin, Y. (1988) Morphology, development and evolutionary significance of the working stages in the caste system of Prorhinotermes (Insecta, Isoptera). Zoomorphology, 107, 339–347.

Romaniuk, A. , Paszel‐Jaworska, A. , Totoń, E. , Lisiak, N. , Hołysz, H. , Królak, A. et al. (2019) The non‐canonical functions of telomerase: to turn off or not to turn off. Molecular Biology Reports, 46, 1401–1411. PubMed

Saebøe‐Larssen, S. , Fossberg, E. and Gaudernack, G. (2006) Characterization of novel alternative splicing sites in human telomerase reverse transcriptase (hTERT): analysis of expression and mutual correlation in mRNA isoforms from normal and tumour tissues. BMC Molecular Biology, 7, 26. PubMed PMC

Sahin, E. and DePinho, R.A. (2010) Linking functional decline of telomeres, mitochondria and stem cells during ageing. Nature, 464, 520–528. PubMed PMC

Santos, J.H. , Meyer, J.N. , Skorvaga, M. , Annab, L.A. and Van Houten, B. (2004) Mitochondrial hTERT exacerbates free‐radical‐mediated mtDNA damage. Aging Cell, 3, 399–411. PubMed

Saretzki, G. (2014) Extra‐telomeric functions of human telomerase: cancer, mitochondria and oxidative stress. Current Pharmaceutical Design, 20, 6386–6403. PubMed

Sasaki, T. and Fujiwara, H. (2000) Detection and distribution patterns of telomerase activity in insects. European Journal of Biochemistry, 267, 3025–3031. PubMed

Ségal‐Bendirdjian, E. and Geli, V. (2019) Non‐canonical roles of telomerase: unraveling the imbroglio. Frontiers in Cell and Developmental Biology, 7, 332. PubMed PMC

Séité, S. , Harrison, M.C. , Sillam‐Dussès, D. , Lupoli, R. , Van Dooren, T.J.M. , Robert, A. et al. (2022) Lifespan prolonging mechanisms and insulin upregulation without fat accumulation in long‐lived reproductives of a higher termite. Communications Biology, 5, 44. PubMed PMC

Shaheen, F. , Grammatopoulos, D.K. , Müller, J. , Zammit, V.A. and Lehnert, H. (2014) Extra‐nuclear telomerase reverse transcriptase (TERT) regulates glucose transport in skeletal muscle cells. Biochimica et Biophysica Acta et Biophysica Acta, 1842, 1762–1769. PubMed

Smith, S. , Hoelzl, F. , Zahn, S. and Criscuolo, F. (2022) Telomerase activity in ecological studies: what are its consequences for individual physiology and is there evidence for effects and trade‐offs in wild populations. Molecular Ecology, 31, 6239–6251. PubMed PMC

Spilsbury, A. , Miwa, S. , Attems, J. and Saretzki, G. (2015) The role of telomerase protein TERT in Alzheimer's disease and in tau‐related pathology in vitro . Journal of Neuroscience, 35, 1659–1674. PubMed PMC

Sun, P. , Li, G. , Jian, J. , Liu, L. , Chen, J. , Yu, S. et al. (2019) Transcriptomic and functional analyses of phenotypic plasticity in a higher termite, Macrotermes barneyi light. Frontiers in Genetics, 10, 964. PubMed PMC

Tasaki, E. , Kobayashi, K. , Matsuura, K. and Iuchi, Y. (2017) An efficient antioxidant system in a long‐lived termite queen. PLoS ONE, 12, e0167412. PubMed PMC

Tasaki, E. , Kobayashi, K. , Matsuura, K. and Yoshihito, I. (2019) Long‐lived termite queens exhibit high Cu/Zn‐superoxide dismutase activity. Oxidative Medicine and Cellular Longevity, 2018, 5127251. PubMed PMC

Tasaki, E. , Takata, M. and Matsuura, K. (2021) Why and how do termite kings and queens live so long? Philosophical Transactions of the Royal Society B ‐ Biological Sciences, 376, 20190740. PubMed PMC

Tasaki, E. , Yamamoto, Y. and Iuchi, Y. (2023) Higher levels of the lipophilic antioxidants coenzyme Q10 and vitamin E in long‐lived termite queens than in short‐lived workers. Insect Science, 31, 201–210. PubMed

Ulaner, G.A. , Hu, J.F. , Vu, T.H. , Giudice, L.C. and Hoffman, A.R. (1998) Telomerase activity in human development is regulated by human telomerase reverse transcriptase (hTERT) transcription and by alternate splicing of hTERT transcripts. Cancer Research, 58, 4168–4172. PubMed

Ulaner, G.A. , Hu, J.F. , Vu, T.H. , Giudice, L.C. and Hoffman, A.R. (2001) Tissue‐specific alternate splicing of human telomerase reverse transcriptase (hTERT) influences telomere lengths during human development. International Journal of Cancer, 91, 644–649. PubMed

Wardi, L. , Alaaeddine, N. , Raad, I. , Sarkis, R. , Serhal, R. , Khalil, C. et al. (2014) Glucose restriction decreases telomerase activity and enhances its inhibitor response on breast cancer cells: possible extra‐telomerase role of BIBR 1532. Cancer Cell International, 14, 60. PubMed PMC

Whittemore, K. , Vera, E. , Martínez‐Nevado, E. , Sanpera, C. and Blasco, M.A. (2019) Telomere shortening rate predicts species life span. Proceedings of the National Academy of Sciences USA, 116, 15122–15127. PubMed PMC

Withers, J.B. , Ashvetiya, T. and Beemon, K.L. (2012) Exclusion of exon 2 is a common mRNA splice variant of primate telomerase reverse transcriptases. PLoS ONE, 7, e48016. PubMed PMC

Ye, C. , Rasheed, H. , Ran, Y. , Yang, X. , Xing, L. and Su, X. (2019) Transcriptome changes reveal the genetic mechanisms of the reproductive plasticity of workers in lower termites. BMC Genomics, 20, 702. PubMed PMC

Yi, X. , Shay, J.W. and Wright, W.E. (2001) Quantitation of telomerase components and hTERT mRNA splicing patterns in immortal human cells. Nucleic Acids Research, 29, 4818–4825. PubMed PMC

Yi, X. , Tesmer, V.M. , Savre‐Train, I. , Shay, J.W. and Wright, W.E. (1999) Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. Molecular and Cellular Biology, 19, 3989–3997. PubMed PMC

Yi, X. , White, D.M. , Aisner, D.L. , Baur, J.A. , Wright, W.E. and Shay, J.W. (2000) An alternate splicing variant of the human telomerase catalytic subunit inhibits telomerase activity. Neoplasia, 2, 433–440. PubMed PMC

Zheng, Q. , Huang, J. and Wang, G. (2019) Mitochondria, telomeres and telomerase subunits. Frontiers in Cell and Developmental Biology, 7, 274. PubMed PMC

Zhu, S. , Rousseau, P. , Lauzon, C. , Gandin, V. , Topisirovic, I. and Autexier, C. (2014) Inactive C‐terminal telomerase reverse transcriptase insertion splicing variants are dominant‐negative inhibitors of telomerase. Biochimie, 101, 93–103. PubMed