Cell Stress and Chaperones

, Volume 18, Issue 3, pp 307–319 | Cite as

A modified UPR stress sensing system reveals a novel tissue distribution of IRE1/XBP1 activity during normal Drosophila development

  • Michio Sone
  • Xiaomei Zeng
  • Joseph Larese
  • Hyung Don Ryoo
Original Paper


Eukaryotic cells respond to stress caused by the accumulation of unfolded/misfolded proteins in the endoplasmic reticulum by activating the intracellular signaling pathways referred to as the unfolded protein response (UPR). In metazoans, UPR consists of three parallel branches, each characterized by its stress sensor protein, IRE1, ATF6, and PERK, respectively. In Drosophila, IRE1/XBP1 pathway is considered to function as a major branch of UPR; however, its physiological roles during the normal development and homeostasis remain poorly understood. To visualize IRE1/XBP1 activity in fly tissues under normal physiological conditions, we modified previously reported XBP1 stress sensing systems (Souid et al., Dev Genes Evol 217: 159–167, 2007; Ryoo et al., EMBO J 26: 242-252, 2007), based on the recent reports regarding the unconventional splicing of XBP1/HAC1 mRNA (Aragon et al., Nature 457: 736–740, 2009; Yanagitani et al., Mol Cell 34: 191–200, 2009; Science 331: 586–589, 2011). The improved XBP1 stress sensing system allowed us to detect new IRE1/XBP1 activities in the brain, gut, Malpighian tubules, and trachea of third instar larvae and in the adult male reproductive organ. Specifically, in the larval brain, IRE1/XBP1 activity was detected exclusively in glia, although previous reports have largely focused on IRE1/XBP1 activity in neurons. Unexpected glial IRE1/XBP1 activity may provide us with novel insights into the brain homeostasis regulated by the UPR.


UPR ER stress XBP1 Drosophila Glia 



The authors thank Professor Toshiya Endo (Nagoya University) and Professor Shuh-ichi Nishikawa (Niigata University) for their helpful encouragement throughout this study. This work was supported by grants from the NEI (R01EY020866) and the Ellison Medical Foundation to H.D.R. and an NIDDK training grant to J.L. (5T35DK007421).


  1. Ali MM, Bagratuni T, Davenport EL, Nowak PR, Silva-Santisteban MC, Hardcastle A, McAndrews C, Rowlands MG, Morgan GJ, Aherne W, Collins I, Davies FE, Pearl LH (2011) Structure of the Ire1 autophosphorylation complex and implications for the unfolded protein response. EMBO J 30:894–905PubMedCrossRefGoogle Scholar
  2. Aragon T, van Anken E, Pincus D, Serafimova IM, Korennykh AV, Rubio CA, Walter P (2009) Messenger RNA targeting to endoplasmic reticulum stress signaling sites. Nature 457:736–740PubMedCrossRefGoogle Scholar
  3. Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415PubMedGoogle Scholar
  4. Casali A, Batlle E (2009) Intestinal stem cells in mammals and Drosophila. Cell Stem Cell 4:124–127PubMedCrossRefGoogle Scholar
  5. Chapman T (2001) Seminal fluid-mediated fitness traits in Drosophila. Heredity 87:511–521PubMedCrossRefGoogle Scholar
  6. Chawla A, Chakrabarti S, Ghosh G, Niwa M (2011) Attenuation of yeast UPR is essential for survival and is mediated by IRE1 kinase. J Cell Biol 193:41–50PubMedCrossRefGoogle Scholar
  7. Cox JS, Shamu CE, Walter P (1993) Transcriptional induction of genes encoding endoplasmic reticulum resident proteins requires a transmembrane protein kinase. Cell 73:1197–1206PubMedCrossRefGoogle Scholar
  8. Cunard R, Sharma K (2011) The endoplasmic reticulum stress response and diabetic kidney disease. Am J Physiol Renal Physiol 300:1054–1061CrossRefGoogle Scholar
  9. Dubchak I, Brudno M, Loots GG, Mayor C, Pachter L, Rubin EM, Frazer KA (2000) Active conservation of noncoding sequences revealed by 3-way species comparisons. Genome Res 10:1304–1306PubMedCrossRefGoogle Scholar
  10. Frazer KA et al (2004) VISTA: computational tools for comparative genomics. Nucleic Acids Res 32:W273–W279 (Web Server issue)PubMedCrossRefGoogle Scholar
  11. Freeman MR, Doherty J (2005) Glial cell biology in Drosophila and vertebrates. Trends Neurosci 29:82–90CrossRefGoogle Scholar
  12. Hetz C (2012) The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Mol Cell Biol 13:89–102Google Scholar
  13. Iwawaki T, Akai R, Kohno K, Miura M (2004) A transgenic mouse model for monitoring endoplasmic reticulum stress. Nat Med 10:98–102PubMedCrossRefGoogle Scholar
  14. Kalb JM, Dibenedetto AJ, Wolfner MF (1993) Probing the function Drosophila melanogaster accessory glands by directed cell ablation. Proc Natl Acad Sci 90:8093–8097PubMedCrossRefGoogle Scholar
  15. Kaser A, Lee AH, Franke A, Glickman JN, Zeissig S, Tilg H, Nieuwenhuis EE, Higgins DE, Schreiber S, Glimcher LH, Blumberg RS (2008) XBP1 links ER stress to intestinal inflammation and confers genetic risk for human inflammatory bowel disease. Cell 134:743–756PubMedCrossRefGoogle Scholar
  16. Korennykh AV, Egea PF, Korostelev AA, Finer-Moore J, Zhang C, Shokat KM, Stroud RM, Walter P (2009) The unfolded protein response signals through high-order assembly of Ire1. Nature 457:687–693PubMedCrossRefGoogle Scholar
  17. Korennykh AV, Korostelev AA, Egea PF, Finer-Moore J, Stroud RM, Zhang C, Shokat KM, Walter P (2011a) Structural and functional basis for RNA cleavage by Ire1. BMC Biol 9:47PubMedCrossRefGoogle Scholar
  18. Korennykh AV, Korostelev AA, Egea PF, Finer-Moore J, Stroud RM, Zhang C, Shokat KM, Walter P (2011b) Cofactor-mediated conformational control in the bifunctional kinase/RNase Ire1. BMC Biol 9:48PubMedCrossRefGoogle Scholar
  19. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132PubMedCrossRefGoogle Scholar
  20. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  21. Lee KP, Dey M, Neculai D, Cao C, Dever TE, Sicheri F (2008a) Structure of the dual enzyme Ire1 reveals the basis for catalysis and regulation in nonconventional RNA splicing. Cell 132:89–100PubMedCrossRefGoogle Scholar
  22. Lee AH, Scapa EF, Cohen DE, Glimcher LH (2008b) Regulation of hepatic lipogenesis by the transcription factor XBP1. Science 320:1492–1496PubMedCrossRefGoogle Scholar
  23. Lin W, Popko B (2009) Endoplasmic reticulum stress in disorders of myelinationg cells. Nature Neurosci 12:379–385PubMedCrossRefGoogle Scholar
  24. Mori K, Ma W, Gething MJ, Sambrook J (1993) A transmembrane protein with a cdc2+/CDC28- related kinase activity is required for signaling from the ER to the nucleus. Cell 74:743–756PubMedCrossRefGoogle Scholar
  25. Parmar VM, Schröder M (2012) Sensing endoplasmic reticulum stress. Adv Exp Med Biol 738:153–168PubMedCrossRefGoogle Scholar
  26. Pennuto M et al (2008) Ablation of the UPR-mediator CHOP restores motor function and reduces demyelination in Charcot-Marie-Tooth 1B mice. Neuron 57:393–405PubMedCrossRefGoogle Scholar
  27. Pereanu W, Spindler S, Cruz L, Hartenstein V (2007) Tracheal development in the Drosophila brain is constrained by glial cells. Dev Biol 302:169–180PubMedCrossRefGoogle Scholar
  28. Richardson CE, Kooistra T, Kim DH (2010) An essential role for XBP-1 in host protection against immune activation in C. elegans. Nature 463:1092–1095PubMedCrossRefGoogle Scholar
  29. Ron D, Ito K (2011) A translational pause to localize. Science 331:543–544PubMedCrossRefGoogle Scholar
  30. Rubio C, Pincus D, Korennykh A, Schuck S, El-Samad H, Walter P (2011) Homeostatic adaptation to endoplasmic reticulum stress depends on Ire1 kinase activity. J Cell Biol 193:171–184PubMedCrossRefGoogle Scholar
  31. Ryoo HD, Steller H (2007) Unfolded protein response in Drosophila: why another model can make it fly. Cell Cycle 6(7):830–835PubMedCrossRefGoogle Scholar
  32. Ryoo HD, Domingos PM, Kang MJ, Steller H (2007) Unfolded protein response in a Drosophila model for retinal degeneration. EMBO J 26:242–252PubMedCrossRefGoogle Scholar
  33. Shen X, Ellis RE, Lee K, Liu CY, Yang Q, Solomon A, Yoshida H, Morimoto R, Kurnit DM, Mori K, Kaufman RJ (2001) Complementary signaling pathways regulate the unfolded protein response and are required for C. elegans development. Cell 107:893–903PubMedCrossRefGoogle Scholar
  34. Shim J, Umemura T, Nothstein E, Rongo C (2004) The unfolded protein response regulates glutamate receptor export from the endoplasmic reticulum. Mol Biol Cell 15:4818–4828PubMedCrossRefGoogle Scholar
  35. Souid S, Lepesant JA, Yanicostas C (2007) The xbp-1 gene is essential for development in Drosophila. Dev Genes Evol 217:159–167PubMedCrossRefGoogle Scholar
  36. Stork T, Bernardos R, Freeman MR (2010) Analysis of glial cell development and function in Drosophila. In: Zhang B, Freeman MR, Waddell S (eds) Drosophila neurobiology: a laboratory manual. Cold Spring Harbor Laboratory Press, New York, pp 53–74Google Scholar
  37. Tsarouhas V, Senti KA, Jayaram SA, Tiklova K, Hemphala J, Adler J, Samakovlis C (2007) Sequential pulses of apical epithelial secretion and endocytosis drive airway maturation in Drosophila. Dev Cell 13:214–225PubMedCrossRefGoogle Scholar
  38. Walter P, Ron D (2011) The unfolded protein response: from stress pathway to homeostatic regulation. Science 334:1081–1086PubMedCrossRefGoogle Scholar
  39. Wiseman RL, Zhang Y, Lee KPK, Harding HP, Haynes CM, Price J, Sicheri F, Ron D (2010) Flavonol activation defines an unanticipated ligand binding site in the kinase-rnase domain of IRE11. Mol Cell 38:291–304PubMedCrossRefGoogle Scholar
  40. Wolfner MF (1997) Tokens of love: functions and regulation of Drosophila male accessory gland products. Insect Biochem Mol Biol 27:179–192PubMedCrossRefGoogle Scholar
  41. Yamamoto K, Sato T, Matsui T, Sato M, Okada T, Yoshida H, Harada A, Mori K (2007) Transcriptional induction of mammalian ER quality control proteins is mediated by single or combined action of ATF6alpha and XBP1. Dev Cell 13:365–376PubMedCrossRefGoogle Scholar
  42. Yanagitani K, Imagawa Y, Iwawaki T, Hosoda A, Saito M, Kimata Y, Kohno K (2009) Cotranslational targeting of XBP1 protein to the membrane promotes cytoplasmic splicing of its own mRNA. Mol Cell 34:191–200PubMedCrossRefGoogle Scholar
  43. Yanagitani K, Kimata Y, Kadokura H, Kohno K (2011) Translational pausing ensures membrane targeting and cytoplasmic splicing of XBP1u mRNA. Science 331:586–589PubMedCrossRefGoogle Scholar
  44. Yoshida H, Oku M, Suzuki M, Mori K (2006) pXBP1(U) encoded in XBP1 pre-mRNA negatively regulates unfolded protein response activator pXBP1(S) in mammalian ER stress response. J Cell Biol 172:565–575PubMedCrossRefGoogle Scholar

Copyright information

© Cell Stress Society International 2012

Authors and Affiliations

  • Michio Sone
    • 1
  • Xiaomei Zeng
    • 1
  • Joseph Larese
    • 1
  • Hyung Don Ryoo
    • 1
  1. 1.Department of Cell BiologyNew York University School of MedicineNew YorkUSA

Personalised recommendations