科技科学

Lsp2 links mTORC1 to TOP mRNA translation and lifespan in <i>Drosophila</i>

Nature作者：Jun Wang2026年9月23日正文已收录本站

Data availability

All relevant datasets generated in this work are publicly available. Regular RNA sequencing data have been deposited in the Gene Expression Omnibus (GEO) under the accession number GSE295794. Translatomic data are available through the GEO under the accession number GSE297586. The proteomic data have been deposited to the ProteomeXchange Consortium (https://proteomecentral.proteomexchange.org) via the iProX partner repository with the identifier PXD063648. Other publicly available databases and datasets that were used in this work: the D. melanogaster protein database (https://www.uniprot.org/taxonomy/7227), the D. melanogaster (BDGP6.46) reference genome (https://www.ensembl.org/Drosophila_melanogaster), FlyAtlas 2 (https://flyatlas.gla.ac.uk/FlyAtlas2/), the KEGG database (https://www.genome.jp/kegg/) and a total of 49 CAGE-seq datasets for 7 species (complete accession numbers are provided in Supplementary Data 8), including the following datasets used for TOP mRNA prediction in Drosophila: SRR27927240, SRR27927241, SRR27927242, SRR488271, SRR488279 and SRR488280 (refs. 72,73). Source data are provided with this paper.

Code availability

Original code for TOP mRNA, mRNA sequencing, monosome sequencing and polysome sequencing data analyses is publicly available via Zenodo at https://doi.org/10.5281/zenodo.21473950 (ref. 74).

References

  1. Liu, G. Y. & Sabatini, D. M. mTOR at the nexus of nutrition, growth, ageing and disease. Nat. Rev. Mol. Cell Biol. 21, 183–203 https://doi.org/10.1038/s41580-019-0199-y (2020).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  2. Meyuhas, O. & Kahan, T. The race to decipher the top secrets of TOP mRNAs. Biochim. Biophys. Acta 1849, 801–811 https://doi.org/10.1016/j.bbagrm.2014.08.015 (2015).

    Article  PubMed  CAS  Google Scholar 

  3. Thoreen, C. C. et al. A unifying model for mTORC1-mediated regulation of mRNA translation. Nature 485, 109–113 https://doi.org/10.1038/nature11083 (2012).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  4. Valzania, L., Alami, A. & Leopold, P. A temporal allocation of amino acid resources ensures fitness and body allometry in Drosophila. Dev. Cell 59, 2277–2286 https://doi.org/10.1016/j.devcel.2024.05.018 (2024).

    Article  PubMed  CAS  Google Scholar 

  5. Fontana, L. & Partridge, L. Promoting health and longevity through diet: from model organisms to humans. Cell 161, 106–118 https://doi.org/10.1016/j.cell.2015.02.020 (2015).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  6. Obata, F. & Miura, M. Regulatory mechanisms of aging through the nutritional and metabolic control of amino acid signaling in model organisms. Annu. Rev. Genet. 58, 19–41 https://doi.org/10.1146/annurev-genet-111523-102042 (2024).

    Article  PubMed  CAS  Google Scholar 

  7. Valvezan, A. J. & Manning, B. D. Molecular logic of mTORC1 signalling as a metabolic rheostat. Nat. Metab. 1, 321–333 https://doi.org/10.1038/s42255-019-0038-7 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. MacInnes, A. W. The role of the ribosome in the regulation of longevity and lifespan extension. Wiley Interdiscip. Rev. RNA 7, 198–212 https://doi.org/10.1002/wrna.1325 (2016).

    Article  PubMed  CAS  Google Scholar 

  9. Bjedov, I. et al. Mechanisms of life span extension by rapamycin in the fruit fly Drosophila melanogaster. Cell Metab. 11, 35–46 https://doi.org/10.1016/j.cmet.2009.11.010 (2010).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Johnson, S. C., Rabinovitch, P. S. & Kaeberlein, M. mTOR is a key modulator of ageing and age-related disease. Nature 493, 338–345 https://doi.org/10.1038/nature11861 (2013).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  11. Mannick, J. B. & Lamming, D. W. Targeting the biology of aging with mTOR inhibitors. Nat. Aging 3, 642–660 https://doi.org/10.1038/s43587-023-00416-y (2023).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Hsieh, A. C. et al. The translational landscape of mTOR signalling steers cancer initiation and metastasis. Nature 485, 55–61 https://doi.org/10.1038/nature10912 (2012).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  13. Peter, D. et al. Molecular architecture of 4E-BP translational inhibitors bound to eIF4E. Mol. Cell 57, 1074–1087 https://doi.org/10.1016/j.molcel.2015.01.017 (2015).

    Article  PubMed  CAS  Google Scholar 

  14. Gingras, A. C. et al. Hierarchical phosphorylation of the translation inhibitor 4E-BP1. Genes Dev. 15, 2852–2864 https://doi.org/10.1101/gad.912401 (2001).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Morita, M. et al. mTOR controls mitochondrial dynamics and cell survival via MTFP1. Mol. Cell 67, 922–935 https://doi.org/10.1016/j.molcel.2017.08.013 (2017).

    Article  PubMed  CAS  Google Scholar 

  16. Tatar, M., Post, S. & Yu, K. Nutrient control of Drosophila longevity. Trends Endocrinol. Metab. 25, 509–517 https://doi.org/10.1016/j.tem.2014.02.006 (2014).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Teleman, A. A., Chen, Y. W. & Cohen, S. M. 4E-BP functions as a metabolic brake used under stress conditions but not during normal growth. Genes Dev. 19, 1844–1848 https://doi.org/10.1101/gad.341505 (2005).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Tettweiler, G., Miron, M., Jenkins, M., Sonenberg, N. & Lasko, P. F. Starvation and oxidative stress resistance in Drosophila are mediated through the eIF4E-binding protein, d4E-BP. Genes Dev. 19, 1840–1843 https://doi.org/10.1101/gad.1311805 (2005).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Zid, B. M. et al. 4E-BP extends lifespan upon dietary restriction by enhancing mitochondrial activity in Drosophila. Cell 139, 149–160 https://doi.org/10.1016/j.cell.2009.07.034 (2009).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Bai, H., Post, S., Kang, P. & Tatar, M. Drosophila longevity assurance conferred by reduced insulin receptor substrate chicopartially requires d4eBP. PLoS ONE 10, e0134415 https://doi.org/10.1371/journal.pone.0134415 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Grandison, R. C., Piper, M. D. & Partridge, L. Amino-acid imbalance explains extension of lifespan by dietary restriction in Drosophila. Nature 462, 1061–1064 https://doi.org/10.1038/nature08619 (2009).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  22. Beneš, H. et al. Adult expression of the Drosophila Lsp-2 gene. Dev. Biol. 142, 138–146 https://doi.org/10.1016/0012-1606(90)90157-e (1990).

    Article  PubMed  Google Scholar 

  23. Sang, J. H. & King, R. C. Nutritional requirements of axenically cultured Drosophila melanogaster adults. J. Exp. Biol. 38, 793–809 https://doi.org/10.1242/jeb.38.4.793 (1961).

    Article  CAS  Google Scholar 

  24. Tiebe, M. et al. REPTOR and REPTOR-BP regulate organismal metabolism and transcription downstream of TORC1. Dev. Cell 33, 272–284 https://doi.org/10.1016/j.devcel.2015.03.013 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Liu, G. Y., Jouandin, P., Bahng, R. E., Perrimon, N. & Sabatini, D. M. An evolutionary mechanism to assimilate new nutrient sensors into the mTORC1 pathway. Nat. Commun. 15, 2517 https://doi.org/10.1038/s41467-024-46680-3 (2024).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  26. Gu, X. et al. Sestrin mediates detection of and adaptation to low-leucine diets in Drosophila. Nature 608, 209–216 https://doi.org/10.1038/s41586-022-04960-2 (2022).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  27. Lu, J. et al. Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids. Nat. Aging 1, 60–72 https://doi.org/10.1038/s43587-020-00001-7 (2021).

    Article  PubMed  Google Scholar 

  28. Hwangbo, D. S., Gershman, B., Tu, M. P., Palmer, M. & Tatar, M. Drosophila dFOXO controls lifespan and regulates insulin signalling in brain and fat body. Nature 429, 562–566 https://doi.org/10.1038/nature02549 (2004).

    Article  ADS  PubMed  CAS  Google Scholar 

  29. Figlia, G. et al. mTORC1 senses glutamine and other amino acids through GCN2. EMBO J. 44, 4825–4866 https://doi.org/10.1038/s44318-025-00505-1 (2025).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. Ye, J. et al. GCN2 sustains mTORC1 suppression upon amino acid deprivation by inducing Sestrin2. Genes Dev. 29, 2331–2336 https://doi.org/10.1101/gad.269324.115 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Carvalho-Santos, Z. & Ribeiro, C. Gonadal ecdysone titers are modulated by protein availability but do not impact protein appetite. J. Insect Physiol. 106, 30–35 https://doi.org/10.1016/j.jinsphys.2017.08.006 (2018).

    Article  PubMed  CAS  Google Scholar 

  32. Benes, H. et al. Overlapping Lsp-2 gene sequences target expression to both the larval and adult Drosophila fat body. Insect Mol. Biol. 5, 39–49 https://doi.org/10.1111/j.1365-2583.1996.tb00039.x (1996).

    Article  PubMed  CAS  Google Scholar 

  33. Kapahi, P. et al. Regulation of lifespan in Drosophila by modulation of genes in the TOR signaling pathway. Curr. Biol. 14, 885–890 https://doi.org/10.1016/j.cub.2004.03.059 (2004).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. Bass, T. M. et al. Optimization of dietary restriction protocols in Drosophila. J. Gerontol. A 62, 1071–1081 https://doi.org/10.1093/gerona/62.10.1071 (2007).

    Article  Google Scholar 

  35. Clancy, D. J. et al. Extension of life-span by loss of CHICO, a Drosophila insulin receptor substrate protein. Science 292, 104–106 https://doi.org/10.1126/science.1057991 (2001).

    Article  ADS  PubMed  CAS  Google Scholar 

  36. Tatar, M. et al. A mutant Drosophila insulin receptor homolog that extends life-span and impairs neuroendocrine function. Science 292, 107–110 https://doi.org/10.1126/science.1057987 (2001).

    Article  ADS  PubMed  CAS  Google Scholar 

  37. O’Brien, D. M., Min, K.-J., Larsen, T. & Tatar, M. Use of stable isotopes to examine how dietary restriction extends Drosophila lifespan. Curr. Biol. 18, R155–R156 https://doi.org/10.1016/j.cub.2008.01.021 (2008).

    Article  PubMed  CAS  Google Scholar 

  38. Yamamoto, R., Palmer, M., Koski, H., Curtis-Joseph, N. & Tatar, M. Aging modulated by the Drosophila insulin receptor through distinct structure-defined mechanisms. Genetics 217, iyaa037 https://doi.org/10.1093/genetics/iyaa037 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  39. Zanco, B., Mirth, C. K., Sgrò, C. M. & Piper, M. D. A dietary sterol trade-off determines lifespan responses to dietary restriction in Drosophila melanogaster females. eLife 10, e62335 https://doi.org/10.7554/eLife.62335 (2021).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Yamamoto, R., Bai, H., Dolezal, A. G., Amdam, G. & Tatar, M. Juvenile hormone regulation of Drosophila aging. BMC Biol. 11, 85 https://doi.org/10.1186/1741-7007-11-85 (2013).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  41. Wek, R. C. Role of eIF2α kinases in translational control and adaptation to cellular stress. Cold Spring Harb. Perspect. Biol. 10, a032870 https://doi.org/10.1101/cshperspect.a032870 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Kim, H. S. et al. Early-adulthood spike in protein translation drives aging via juvenile hormone/germline signaling. Nat. Commun. 14, 5021 https://doi.org/10.1038/s41467-023-40618-x (2023).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  43. Recasens-Alvarez, C. et al. Ribosomopathy-associated mutations cause proteotoxic stress that is alleviated by TOR inhibition. Nat. Cell Biol. 23, 127–135 https://doi.org/10.1038/s41556-020-00626-1 (2021).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. Sung, M. K. et al. A conserved quality-control pathway that mediates degradation of unassembled ribosomal proteins. eLife 5, e19105 https://doi.org/10.7554/eLife.19105 (2016).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  45. Romero-Pozuelo, J., Demetriades, C., Schroeder, P. & Teleman, A. A. CycD/Cdk4 and discontinuities in Dpp signaling activate TORC1 in the Drosophila wing disc. Dev. Cell 42, 376–387 https://doi.org/10.1016/j.devcel.2017.07.019 (2017).

    Article  PubMed  CAS  Google Scholar 

  46. Kosakamoto, H. et al. Lsp2 links early-life diet to adult translation and lifespan in Drosophila. Nature https://doi.org/10.1038/s41586-026-11031-3 (2026).

  47. Philippe, L., van den Elzen, A. M. G., Watson, M. J. & Thoreen, C. C. Global analysis of LARP1 translation targets reveals tunable and dynamic features of 5′ TOP motifs. Proc. Natl Acad. Sci. USA 117, 5319–5328 https://doi.org/10.1073/pnas.1912864117 (2020).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  48. Meyuhas, O. Synthesis of the translational apparatus is regulated at the translational level. Eur. J. Biochem. 267, 6321–6330 https://doi.org/10.1046/j.1432-1327.2000.01719.x (2000).

    Article  PubMed  CAS  Google Scholar 

  49. Marygold, S. J. et al. The ribosomal protein genes and Minute loci of Drosophila melanogaster. Genome Biol. 8, R216 https://doi.org/10.1186/gb-2007-8-10-r216 (2007).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Leader, D. P., Krause, S. A., Pandit, A., Davies, S. A. & Dow, J. A. T. FlyAtlas 2: a new version of the Drosophila melanogaster expression atlas with RNA-seq, miRNA-seq and sex-specific data. Nucleic Acids Res. 46, D809–D815 https://doi.org/10.1093/nar/gkx976 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Hernández, G., Altmann, M. & Lasko, P. Origins and evolution of the mechanisms regulating translation initiation in eukaryotes. Trends Biochem. Sci. 35, 63–73 https://doi.org/10.1016/j.tibs.2009.10.009 (2010).

    Article  PubMed  CAS  Google Scholar 

  52. Sun, J. et al. Drosophila FIT is a protein-specific satiety hormone essential for feeding control. Nat. Commun. 8, 14161 https://doi.org/10.1038/ncomms14161 (2017).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  53. Vidarsson, G., Dekkers, G. & Rispens, T. IgG subclasses and allotypes: from structure to effector functions. Front. Immunol. 5, 520 https://doi.org/10.3389/fimmu.2014.00520 (2014).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  54. Jones, D. D. et al. mTOR has distinct functions in generating versus sustaining humoral immunity. J. Clin. Invest. 126, 4250–4261 https://doi.org/10.1172/JCI86504 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  55. Yu, L. et al. IgG is an aging factor that drives adipose tissue fibrosis and metabolic decline. Cell Metab. 36, 793–807 https://doi.org/10.1016/j.cmet.2024.01.015 (2024).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  56. Ma, S. et al. Spatial transcriptomic landscape unveils immunoglobin-associated senescence as a hallmark of aging. Cell 187, 7025–7044 https://doi.org/10.1016/j.cell.2024.10.019 (2024).

    Article  PubMed  CAS  Google Scholar 

  57. Jin, H. et al. TRIBE editing reveals specific mRNA targets of eIF4E-BP in Drosophila and in mammals. Sci. Adv. 6, eabb8771 https://doi.org/10.1126/sciadv.abb8771 (2020).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  58. Berman, A. J. et al. Controversies around the function of LARP1. RNA Biol. 18, 207–217 https://doi.org/10.1080/15476286.2020.1733787 (2021).

    Article  PubMed  CAS  Google Scholar 

  59. Blagden, S. P. et al. Drosophila Larp associates with poly(A)-binding protein and is required for male fertility and syncytial embryo development. Dev. Biol. 334, 186–197 https://doi.org/10.1016/j.ydbio.2009.07.016 (2009).

    Article  PubMed  CAS  Google Scholar 

  60. Narla, A. & Ebert, B. L. Ribosomopathies: human disorders of ribosome dysfunction. Blood 115, 3196–3205 https://doi.org/10.1182/blood-2009-10-178129 (2010).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  61. McGuire, S. E., Mao, Z. & Davis, R. L. Spatiotemporal gene expression targeting with the TARGET and gene-switch systems in Drosophila. Sci. STKE 2004, pl6 https://doi.org/10.1126/stke.2202004pl6 (2004).

    Article  PubMed  Google Scholar 

  62. Osterwalder, T., Yoon, K. S., White, B. H. & Keshishian, H. A conditional tissue-specific transgene expression system using inducible GAL4. Proc. Natl Acad. Sci. USA 98, 12596–12601 https://doi.org/10.1073/pnas.221303298 (2001).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  63. Roman, G., Endo, K., Zong, L. & Davis, R. L. P{Switch}, a system for spatial and temporal control of gene expression in Drosophila melanogaster. Proc. Natl Acad. Sci. USA 98, 12602–12607 https://doi.org/10.1073/pnas.221303998 (2001).

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  64. Zhang, S. et al. Eclosion muscles secrete ecdysteroids to initiate asymmetric intestinal stem cell division in Drosophila. Dev. Cell 59, 125–140 https://doi.org/10.1016/j.devcel.2023.11.016 (2024).

    Article  PubMed  CAS  Google Scholar 

  65. Port, F. et al. A large-scale resource for tissue-specific CRISPR mutagenesis in Drosophila. eLife 9, e53865 https://doi.org/10.7554/eLife.53865 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  66. Piper, M. D. et al. A holidic medium for Drosophila melanogaster. Nat. Methods 11, 100–105 https://doi.org/10.1038/nmeth.2731 (2014).

    Article  PubMed  CAS  Google Scholar 

  67. Kosakamoto, H. et al. Sensing of the non-essential amino acid tyrosine governs the response to protein restriction in Drosophila. Nat. Metab. 4, 944–959 https://doi.org/10.1038/s42255-022-00608-7 (2022).

    Article  PubMed  CAS  Google Scholar 

  68. Zhai, Z., Boquete, J. P. & Lemaitre, B. Cell-specific Imd-NF-κB responses enable simultaneous antibacterial immunity and intestinal epithelial cell shedding upon bacterial infection. Immunity 48, 897–910 https://doi.org/10.1016/j.immuni.2018.04.010 (2018).

    Article  PubMed  CAS  Google Scholar 

  69. Luo, S. et al. Drosophila ts RNAs preferentially suppress general translation machinery via antisense pairing and participate in cellular starvation response. Nucleic Acids Res. 46, 5250–5268 https://doi.org/10.1093/nar/gky189 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  70. Chen, T. et al. The Drosophila NPY-like system protects against chronic stress-induced learning deficit by preventing the disruption of autophagic flux. Proc. Natl Acad. Sci. USA 120, e2307632120 https://doi.org/10.1073/pnas.2307632120 (2023).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  71. Chothani, S. et al. deltaTE: detection of translationally regulated genes by integrative analysis of Ribo-seq and RNA-seq data. Curr Protoc Mol Biol 129, e108 https://doi.org/10.1002/cpmb.108 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  72. Chen, J. et al. Piwi regulates the usage of alternative transcription start sites in the Drosophila ovary. Nucleic Acids Res. 53, gkae1160 https://doi.org/10.1093/nar/gkae1160 (2025).

    Article  PubMed  PubMed Central  Google Scholar 

  73. Hoskins, R. A. et al. Genome-wide analysis of promoter architecture in Drosophila melanogaster. Genome Res. 21, 182–192 https://doi.org/10.1101/gr.112466.110 (2011).

    Article  PubMed  CAS  Google Scholar 

  74. Cai, Z. Code release for “Lsp2 links mTORC1 to TOP mRNA translation and lifespan in Drosophila”. Zenodo https://doi.org/10.5281/zenodo.21473950 (2026).

Download references

Acknowledgements

We thank C. Neyen, M. Soller, S. Sorge, Z. Guo and F. Obata for comments; A. Teleman, Y. Wei, B. Lemaitre, J. Veenstra, J. Chen and Y. Li for sharing flies, antibodies and bacterial strains; the Bloomington Drosophila Stock Center (BDSC), Vienna Drosophila Resource Center (VDRC), TsingHua Fly Center (THFC) and Kyoto Stock Center for fly stocks; DSHB for antibodies; and J. Su, X. Wang and X. Xie for technical support.

Funding

This work was supported by Hunan NSF grants 2025JJ30009 (to Z.Z.) and 2025JJ40028 (to Y. Wang); the Key Project of Developmental Biology and Breeding from Hunan Province (2026XKQ2303 to Z.Z.); NSFC grants 32170509 (to Z.Z.), 32470655 (to Y. Wang) and 32371219 (to J.L.); the Shanghai Pujiang Program (23PJ1415500 to J.L.); and the Hong Kong Research Grants Council (RGC) (GRF16103620 and GRF16104324 to Y.Y.).

Author information

Author notes

  1. These authors contributed equally: Jun Wang, Zixin Cai

Authors and Affiliations

  1. Hunan Research Center of the Basic Discipline for Developmental Biology, College of Life Sciences, Hunan Normal University, Changsha, China

    Jun Wang  (王骏), Jiaojiao Gu  (谷姣姣), Shiqi Xiong  (熊诗琪), Jianhan Yi  (易翦涵), Meng Yang  (杨猛), Kexin Chang  (常可忻) & Zongzhao Zhai  (翟宗昭)

  2. School of Basic Medical Sciences, Hunan Normal University, Changsha, China

    Jun Wang  (王骏)

  3. Hunan Research Center of the Basic Discipline for Cell Signaling, College of Biology, Hunan University, Changsha, China

    Zixin Cai  (蔡子欣), Xinrui Ning  (宁鑫蕊), Yilin Wen  (闻溢琳) & Yirong Wang  (王奕蓉)

  4. Division of Life Science, Hong Kong University of Science and Technology, Hong Kong, China

    Yan Yan  (阎言)

  5. Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, Shanghai, China

    Jiongming Lu  (陆炯明)

Authors

  1. Jun Wang  (王骏)
  2. Zixin Cai  (蔡子欣)
  3. Jiaojiao Gu  (谷姣姣)
  4. Shiqi Xiong  (熊诗琪)
  5. Jianhan Yi  (易翦涵)
  6. Meng Yang  (杨猛)
  7. Kexin Chang  (常可忻)
  8. Xinrui Ning  (宁鑫蕊)
  9. Yilin Wen  (闻溢琳)
  10. Yan Yan  (阎言)
  11. Jiongming Lu  (陆炯明)
  12. Yirong Wang  (王奕蓉)
  13. Zongzhao Zhai  (翟宗昭)

Contributions

J.W. and Z.Z. developed the research plan and, together with J.L. and Y. Wang, interpreted the results. J.W. designed and performed all of the fly work. Z.C. performed polysome profiling and associated RNA sequencing and TOP motif analysis. J.G., S.X., J.Y. and K.C. helped with quantitative PCR, lifespan, infection and food intake analyses. M.Y. helped analyse regular RNA sequencing data. X.N. and Y. Wen helped with polysome profiling. Y.Y. provided a key reagent. J.L., Y. Wang and Z.Z. co-supervised the project. Z.Z. wrote the manuscript and, together with J.L. and Y. Wang, revised it. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Jiongming Lu  (陆炯明), Yirong Wang  (王奕蓉) or Zongzhao Zhai  (翟宗昭).

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature thanks Linda Partridge, Marc Tatar and the other, anonymous reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended Data Fig. 1 Lsp2 is induced by dietary EAAs.

a, Gene ontology (GO) analyses of significantly differentially expressed genes (DEGs, Q < 0.05 & |log2FoldChange|>1) in the YE vs S comparison (Diet), the Old vs Young comparison (Age) and the 309 co-regulated DEGs. Shown are the top 20 significantly enriched GO terms. b, Volcano plots of DEGs with Lsp2 indicated. c, Expression of Drosophila storage protein genes in the transcriptomic datasets. d, Western blots (WBs) showing Lsp2 protein levels in whole flies of 5-day-old w1118 females treated with the indicated diets for 2 days. e, Lsp2 is mainly expressed in the head fat body. Relative Lsp2 mRNA levels (left) in dissected fat body from the heads and from the bodies of 7-day-old w1118 females raised on CD. Lsp2 expression in fly heads indicated by Lsp2-Gal4 > UAS-GFP (right). 5-day-old females were fed 1% S or 1% S with 10% YE for 2 days. f, Relative Lsp2 mRNA levels in heads of 5-day-old mated w1118 females, virgin females, and males raised on the indicated YE diets for 2 days. All diets contained 1% S. g, Relative Lsp2 mRNA levels in heads of 5-day-old mated w1118 females raised on the indicated diets with alternative protein sources for 1 (left, with 1% S) or 2 (right, with 5% S) days. h, Relative Lsp2 mRNA levels in heads of 5-day-old mated w1118 females raised on the indicated AA diets with 1% S for 1 day. AA contents summed to 5% w/v, and each AA was at an equal weight ratio when multiple AAs were included in one diet. BCAA is a mixture of Leu, Ile and Val (middle). In the right panel, the numbers under the x axis stand for no AA supplied (0); Leu, Ile and Val (1, BCAAs); Leu, Ile, Val, Phe and Trp (2, BCAAs + Phe + Trp); Leu, Ile, Val, Phe, Trp, Lys and Arg (3, seven EAAs); Leu, Ile, Val, Phe, Trp, Lys, Arg, Met, Thr and His (4, full EAAs); Phe, Trp, Lys, Arg, Met, Thr and His (5, full EAAs lacking BCAAs); Lys, Arg, Met, Thr and His (6, five EAAs); Met, Thr and His (7, three EAAs). 20 heads/sample per dot in c,f–h. Lsp2 expression on sucrose-only food is normalized to 1 in f–h. Data are mean ± s.e.m. Student’s t-test in c,e; one-way ANOVA with Dunnett’s multiple comparisons test in f–h. All P values shown are from two-sided tests. Shown in d,e are representative images of three biological replicates. Scale bars 50 µm in e.

Source data

Extended Data Fig. 2 Lsp2 expression integrates multiple nutritional and physiological signals.

a, Lsp2 expression is induced by mTORC1 signalling. A schematic of mTORC1 signalling and relative Lsp2 levels in heads of flies with the indicated genotypes. b, Lsp2 expression is activated by insulin signalling. A schematic of insulin signalling and relative Lsp2 levels in heads of flies with the indicated genotypes. yw was used to generate LppTS>Control. c,d, FoxO and ATF4 suppress the induction of Lsp2 expression by EAAs and mTORC1. Relative Lsp2 levels in heads of flies with the indicated genotypes. A schematic of integrated stress response (ISR) signalling is shown in d. e, The UAS lines used have no significant background effect on Lsp2 expression. qPCR detecting relative Lsp2 levels in heads of 7-day-old progeny raised at 25 °C on CD from the indicated crosses. f, A model summarizing the identified factors that gate Lsp2 expression. g, Ecdysone signalling activity in fat body is required for Lsp2 expression. Relative Lsp2 levels in heads of flies expressing Control (from left to right: yw, yw, w1118 and w1118) and the indicated UAS-transgenes alone and in combinations specifically in fat body using LppTS. h, Overexpressing Lsp2 via a UAS-transgene increases the transcription of the endogenous Lsp2 (measured with primers targeting 3′ untranslated region (UTR) of Lsp2). Shown are qPCR assays detecting Lsp2 coding regions (both endogenous and ectopic expression) and 3′ UTR in heads of S32-GS-Gal4 > UAS-Lsp2 (S32>Lsp2) flies with/without the addition of RU486 to induce transgene expression for 3 days on CD at 25 °C. Diet conditions are indicated in figures. Unless otherwise noted, 3-day-old flies were shifted to 29 °C either for 5 days on CD or for 3 days on CD followed by 2 days on control/EAA diets prior to qPCR analysis, and flies for genetic epistatic assays involving FoxO and ATF4 overexpression were shifted to 29 °C for 2 days. 20 female heads/sample per dot in all quantifications. Data are mean ± s.e.m. Student’s t-test for comparison of two groups; two-way ANOVA with Šídák’s multiple comparisons test for experiments with both control and EAA diets; others in one-way ANOVA with Dunnett’s multiple comparisons test. All P values shown are from two-sided tests. Ri, RNAi.

Source data

Extended Data Fig. 3 Lsp2 regulates lifespan.

a,b, Schematic (a) of the generation of Lsp2A10 mutant line and its genetic control Lsp2B14 by mobilizing the indicated P{EP} transposon, and validation of the lines (b) by qPCR and WB. Each dot represents 20 heads of 5-day-old females raised on CD, used for qPCR. 20 whole flies of 5-day-old females raised for 2 days, either on CD or on a diet consisting of 3.3% YE and 2% S, were processed for WB. c, Developmental timing of Lsp2A10 flies and its genetic control on CD. Pupariation time (left) and adult eclosion time (right) are indicated as hours taken after egg-laying (AEL) in a group of about 40 synchronized 1st instar larvae. n = 5 for control; n = 6 for Lsp2A10. d, Lsp2 protein levels in control and Lsp2A10 mutant flies raised on SYA foods of increasing yeast contents for 7 days. Whole flies were used. e, Survival (left) and median lifespan (right) of female Lsp2KO flies (isogenized into wDah background) and control (wDah) on the five SYA diets. Numbers of flies used were 63, 74, 59, 70 and 58 for control; 57, 39, 37, 63 and 53 for Lsp2KO, respectively. The lifespan response to dietary protein levels differed significantly between Lsp2KO and control flies (CPH analysis, P = 0.0004). f–j, Survival analyses of female flies (except right panel in f) of the indicated genotypes raised on the indicated diets. 200 μM RU486 was added from mid-life at day 36 (right panel in i) or from day 10 (right panel in j) to activate transgene expression via the GeneSwitch system. RNAi (Ri) was used to deplete Lsp2 from 10 days of age using either TARGET system (left in j) or GeneSwitch (right in j). Lpp-Gal4 and Cg-Gal4 are pan-adipose drivers; S32-GS drives expression mainly in the head fat body. From f to j, numbers of flies used for control were 134, 87, 125, 102, 114, 122, 91, 120, 129 and 121; for experimental groups, 129, 80, 87, 122, 115, 77, 102, 129, 110 and 114. Data are mean ± s.e.m. in b,c with Student’s t-test. Log-rank test in e–j. All P values shown are from two-sided tests.

Source data

Extended Data Fig. 4 Loss of Lsp2 does not impair fly fitness.

a, Survival of control and Lsp2A10 flies under starvation. 5-day-old flies were analysed on 1% agar tube with only access to water. For female (left), n = 79 in both control and Lsp2A10; for male (right), n = 122 in control and 128 in Lsp2A10. b, Survival of 5-day-old control and Lsp2A10 females to oxidative stress. 17.5 μM paraquat was applied to S5Y10 food. n = 120 for both control and Lsp2A10. c,d, Eggs laid per fly per 24 h (left), climbing distance in the indicated time (left middle), average speed of spontaneous movements monitored for 8 h (right middle) and survival to systemic bacterial infection by Providencia rettgeri (P. rett) (right) in 10-day-old (c) and 30-day-old (d) control and Lsp2A10 females. For egg-laying, each dot represents a group of 10 females and 3 males; for climbing, each dot stands for the average distance climbed by a group of 10 females (6 biological replicates, each performed in technical triplicate); for spontaneous walking, each dot represents one fly; for infection experiments, flies used for control and Lsp2A10 were 28 and 24 (c), and 39 and 39 (d). e,f, Lsp2 is required for timely reproduction but not for fecundity. Monitoring egg-laying of groups of 3 females with 3 males (1-day-old) every 2 days for 30 days for control (n = 15) and Lsp2A10 (n = 17) flies raised on CD (e). Egg-laying of control (n = 17) and Lsp2A10 (n = 17) flies during a fasting-refeeding treatment (f), where groups of 5 females with 3 males (5-day-old) were dry-starved for 6 h and allowed to ingest fresh yeast (FY) for 1 day before being shifted to the S5Y1 diet. Shown are eggs laid each day (left in f) and over 5 days (right in f). Data are mean ± s.e.m. with Student’s t-test except survival with log-rank test. All P values shown are from two-sided tests.

Source data

Extended Data Fig. 5 Lsp2 does not affect insulin and JH activity or global transcription.

a–c, qPCR measuring Drosophila insulin-like peptide 2 (dilp2) mRNA levels (a), ELISA quantification of circulating Dilp2 protein levels (b) and immunofluorescence showing levels of Dilp2 retention in the insulin-producing cells (IPCs) (c) of flies with the indicated genotypes raised on CD, except the control and Lsp2A10 in c on 2% S + 3.3% YE food. Heads (a), haemolymph (b) and brains (c) of 5-day-old females were used. The results show that Lsp2 adipokine acts to limit the release of the brain insulin Dilp2. d, Lsp2 does not affect systemic insulin activity. WBs and quantifications of phosphorylated (p-Akt) and total Akt. Samples used were 7-day-old whole flies of control and Lsp2A10 raised on CD, and 3-day-old LppTS>Control and LppTS>Lsp2 flies that were shifted to 29 °C for 3 days on CD followed by 2 days on 5% S. e, qPCR indicating the JH target gene Kr-h1 is not affected by Lsp2. Samples used were 7-day-old whole flies of control and Lsp2A10 raised on CD. f, RNA-seq analyses suggest Lsp2 is not a transcriptional regulator. PCA analysis (left) of DEGs in heads of adult females with the indicated genotypes raised at 29 °C for 5 days (3 days on CD followed by 2 days on 5% S) to overexpress Lsp2 or Rheb in fat body using LppTS. Very few genes including Hsp70Ba and Hsp70Bb were co-regulated by both Lsp2 and Rheb overexpression. Each dot in a,d,e stands for a group of 20 flies, each dot in b for 50 flies, and each dot in c denotes one brain. Data are mean ± s.e.m in a–e with Student’s t-test. All P values shown are from two-sided tests. Scale bars 50 μm in c.

Source data

Extended Data Fig. 6 Lsp2 regulates RP mRNA translation and lifespan through 4E-BP.

a,b, Enriched KEGG terms in downregulated DEGs in Lsp2A10 flies compared to control (a) and in Thor2; Lsp2A10 flies compared to Thor2; Lsp2B14 flies (b), in levels of mRNA-seq, monosome-seq, polysome-seq and translational efficiency (TE). Shown are the top 5 enriched processes at each level. c,d, Lifespan of control (n = 127) and Lsp2A10 (n = 124) females (c), and Thor2; Lsp2B14 (n = 127) and Thor2; Lsp2A10 (n = 116) females (d) raised on CD. e, Polysome profiling of 15-day-old control and Lsp2A10 flies raised on S5Y5 food reveals an increase in the 60S subunit (top) and a decrease in the 40S subunit (bottom) in Lsp2 mutants. f, Volcano plots of expression changes in mRNA-seq, monosome-seq, polysome-seq, TE, and proteome in Lsp2A10 compared to control flies. Cytoplasmic RP and mitochondrial RP (mRP) mRNAs/proteins are highlighted. The dashed horizontal lines denote thresholds. g, Polysome profiling of 15-day-old Thor2; Lsp2B14 and Thor2; Lsp2A10 flies raised on S5Y5 food reveals similar levels of the 60S subunit (top) and the 40S subunit (bottom). h, Volcano plots of expression changes in mRNA-seq, monosome-seq, polysome-seq, and TE in Thor2; Lsp2A10 double mutant flies compared to Thor2; Lsp2B14 flies. RP and mRP mRNAs are highlighted. The dashed horizontal lines denote thresholds. i, WBs and quantifications of the indicated RP proteins in whole flies of control and Lsp2A10 raised on CD for 7 days. j–l, Loss of Drosophila 4E-BP (Thor2) significantly changes the lifespan response to dietary protein levels (l) without affecting mTORC1 activity (j) or the rate of global protein synthesis (k). We isogenized Thor2 into w1118 iso (iso31) background and used this line with its iso31 control for these experiments. j, WBs and quantifications of phosphorylated (p-S6) and total RpS6 (S6) levels using whole flies of the indicated genotypes raised on CD for 7 days. k, WBs and quantifications of puromycin incorporation in 3-day-old (left) and 15-day-old (right) control and Thor2 flies raised on CD prior to a S5Y10 diet containing 600 μM puromycin for 1 day. Fly heads were used for sequential Ponceau S staining (bottom image) and anti-puromycin detection (top image) for the same gel. l, Survival and median lifespan of control and Thor2 flies raised on SYA foods. From S5Y1 to S5Y20, for control, n = 110, 115, 110 and 113; for Thor2, n = 107, 115, 124, and 117. Each dot stands for a group of 60 flies in e,g and 20 flies/heads in i–k. Data are mean ± s.e.m. in e,g,i–k with Student’s t-test. Log-rank test in c,d,l. All P values shown are from two-sided tests. P stands for polysome and M for monosome in e,g.

Source data

Extended Data Fig. 7 Lsp2 promotes mTORC1 activity in mature adult flies but not during early adulthood.

a,b, Lsp2 fails to increase 4E-BP phosphorylation under conditions of mTORC1 inhibition by expressing mTORDN (a) or mutating Nup44A (a GATOR2 component, b). c, Mutants of GATOR2 components (mio and Nup44A) show reduced mTORC1 activity in adults. WBs and quantifications of Thr37/46-phosphorylated 4E-BP (p-4E-BP), Thr46-dephosphorylated 4E-BP (non-p-4E-BP), phosphorylated RpS6 (p-S6) and total RpS6 (S6) levels using whole flies of the indicated genotypes. 3-day-old flies shifted to 29 °C for 3 days on CD in a,b; 3-day-old flies raised on CD for 5 days at 25 °C in c. mioKO2/+ heterozygous flies were obtained by crossing mioKO2 mutant to yw (control), and Nup44A∆15/+ heterozygous flies were obtained by crossing Nup44A∆15 mutant to w1118 (control). d,e, AA-targeted metabolomics of control and Lsp2A10 flies to measure free AA levels. d, 5-day-old females raised on CD were put on a 5% S + 10% YE diet for 1 day, and then shifted to 5% S food for 0, 2 and 4 days (time points chosen according to Fig. 1f); e, 15-day-old females raised on CD. 40 whole flies/sample in 6 biological replicates. Shown are heat maps indicating the expression levels of each AA. f–h, Lsp2 does not regulate mTORC1 activity or global RP translation in early adulthood. Control and Lsp2A10 flies in otherwise wild-type background or Thor2 background were all used at 3 days after adult emergence on CD. WBs and quantifications measuring p-S6, total S6, p-4E-BP and non-p-4E-BP levels using 40 whole flies/sample (f). Volcano plots of expression changes (top) and enriched KEGG terms (bottom) in downregulated DEGs in mRNA-seq, monosome-seq, polysome-seq and TE in Lsp2A10 compared to control flies (g). Cytosolic RP and mitochondrial RP (mRP) mRNAs are highlighted. The dashed horizontal lines denote thresholds. Note the prominent and global reduction in mRP mRNA translation in Lsp2A10 flies at this early adult stage. WBs and quantifications of puromycin incorporation (h) using 2-day-old flies of the indicated genotypes raised on CD prior to a S5Y10 diet containing 600 μM puromycin for 1 day. 20 fly heads for one sample. Ponceau S staining (bottom image) and anti-puromycin detection (top image) were sequentially performed for the same gel. Data are mean ± s.e.m. in a–c,f,h, in which each dot stands for 20 flies, except in f (40 flies/sample). One-way ANOVA with Dunnett’s multiple comparisons test in a–c; Student’s t-test in f,h. All P values shown are from two-sided tests.

Source data

Extended Data Fig. 8 Long-lived Lsp2A10 flies are susceptible to rapamycin treatment.

a,b, Lifespan analyses of control and Lsp2A10 flies treated with 200 µM rapamycin or its solvent added to CD (a) and S5Y20 food (b). Flies used for control, control + Rapa, Lsp2A10 and Lsp2A10 + Rapa were 76, 94, 79 and 94 in a; 126, 119, 128 and 125 in b. c, A dose–response lifespan assay of control and Lsp2A10 flies treated with rapamycin of increasing concentrations on S5Y10 diet. Shown are survival plots and median lifespan. From low to high rapamycin doses, the numbers of flies used were 128, 127, 120, 126 and 125 for control; 101, 116, 122, 124 and 129 for Lsp2A10. d, Model. Log-rank test in a–c. All P values shown are from two-sided test.

Source data

Extended Data Fig. 9 Stage-specific regulation of RP and TOP mRNA translation.

a, Changes in translational efficiencies (TEs) of the predicted TOP mRNAs (orange bars) compared to that of all mRNAs (grey bars) in the indicated comparisons. Significance was determined by two-sided Mann–Whitney U test. P values indicating differences between changes in TEs of TOP mRNAs and those of all mRNAs are 0.0300, 1.5600 × 10−26 and 2.0996 × 10−9, from left to right graphs. b, Heat maps summarizing changes in TEs of cytosolic RP (left) and mitochondrial RP (mRP, right) mRNAs in the four indicated datasets generated from 15-day-old flies.

Extended Data Table 1 Predicted TOP genes in Drosophila

Full size table

Supplementary information

Source data

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, J., Cai, Z., Gu, J. et al. Lsp2 links mTORC1 to TOP mRNA translation and lifespan in Drosophila. Nature (2026). https://doi.org/10.1038/s41586-026-11029-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • DOI: https://doi.org/10.1038/s41586-026-11029-x