Article | Published:

A CREB3–ARF4 signalling pathway mediates the response to Golgi stress and susceptibility to pathogens

Nature Cell Biology volume 15, pages 14731485 (2013) | Download Citation

Abstract

Treatment of cells with brefeldin A (BFA) blocks secretory vesicle transport and causes a collapse of the Golgi apparatus. To gain more insight into the cellular mechanisms mediating BFA toxicity, we conducted a genome-wide haploid genetic screen that led to the identification of the small G protein ADP-ribosylation factor 4 (ARF4). ARF4 depletion preserves viability, Golgi integrity and cargo trafficking in the presence of BFA, and these effects depend on the guanine nucleotide exchange factor GBF1 and other ARF isoforms including ARF1 and ARF5. ARF4 knockdown cells show increased resistance to several human pathogens including Chlamydia trachomatis and Shigella flexneri. Furthermore, ARF4 expression is induced when cells are exposed to several Golgi-disturbing agents and requires the CREB3 (also known as Luman or LZIP) transcription factor, whose downregulation mimics ARF4 loss. Thus, we have uncovered a CREB3–ARF4 signalling cascade that may be part of a Golgi stress response set in motion by stimuli compromising Golgi capacity.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Ras signalling on the endoplasmic reticulum and the Golgi. Nat. Cell Biol. 4, 343–350 (2002).

  2. 2.

    , , , & A role for Arf1 in mitotic Golgi disassembly, chromosome segregation, and cytokinesis. Proc. Natl Acad. Sci. USA 100, 13314–13319 (2003).

  3. 3.

    , , & Fragmentation and dispersal of the pericentriolar Golgi complex is required for entry into mitosis in mammalian cells. Cell 109, 359–369 (2002).

  4. 4.

    , & A primary role for Golgi positioning in directed secretion, cell polarity, and wound healing. Mol. Biol. Cell 20, 1728–1736 (2009).

  5. 5.

    , & Polarization of the Golgi apparatus and the microtubule-organizing center in cultured fibroblasts at the edge of an experimental wound. Proc. Natl Acad. Sci. USA 79, 2603–2607 (1982).

  6. 6.

    , & Decumbin, a new compound from a species of Penicillium. Nature 181, 1072–1073 (1958).

  7. 7.

    , & Brefeldin A is a potent inducer of apoptosis in human cancer cells independently of p53. Exp. Cell Res. 227, 190–196 (1996).

  8. 8.

    , , , & Induction of terminal differentiation and apoptosis in human colonic carcinoma cells by brefeldin A, a drug affecting ganglioside biosynthesis. FEBS Lett. 453, 140–144 (1999).

  9. 9.

    et al. Antiproliferative effect in vitro and antitumor activity in vivo of brefeldin A. The Cancer J. Sci. Am. 2, 52–58 (1996).

  10. 10.

    , & Synthesis and anticancer activity of brefeldin A ester derivatives. J. Med. Chem. 49, 3897–3905 (2006).

  11. 11.

    & ARF family G proteins and their regulators: roles in membrane transport, development and disease. Nat. Rev. Mol. Cell Biol. 12, 362–375 (2011).

  12. 12.

    & ARF proteins: roles in membrane traffic and beyond. Nat. Rev. Mol. Cell Biol. 7, 347–358 (2006).

  13. 13.

    et al. Brefeldin A acts to stabilize an abortive ARF-GDP-Sec7 domain protein complex: involvement of specific residues of the Sec7 domain. Mol. Cell 3, 275–285 (1999).

  14. 14.

    , , & A regulatory role for ARF6 in receptor-mediated endocytosis. Science 267, 1175–1178 (1995).

  15. 15.

    et al. Overexpression of wild-type and mutant ARF1 and ARF6: distinct perturbations of nonoverlapping membrane compartments. J. Cell Biol. 128, 1003–1017 (1995).

  16. 16.

    et al. Intracellular distribution of Arf proteins in mammalian cells. Arf6 is uniquely localized to the plasma membrane. J. Biol. Chem. 271, 21767–21774 (1996).

  17. 17.

    et al. A haploid genetic screen identifies the major facilitator domain containing 2A (MFSD2A) transporter as a key mediator in the response to tunicamycin. Proc. Natl Acad. Sci. USA 108, 11756–11765 (2011).

  18. 18.

    et al. Haploid genetic screens in human cells identify host factors used by pathogens. Science 326, 1231–1235 (2009).

  19. 19.

    et al. Global gene disruption in human cells to assign genes to phenotypes by deep sequencing. Nature Biotechnol. 29, 542–546 (2011).

  20. 20.

    , , & Isoform-selective effects of the depletion of ADP-ribosylation factors 1-5 on membrane traffic. Mol. Biol. Cell 16, 4495–4508 (2005).

  21. 21.

    et al. ARF1 and ARF4 regulate recycling endosomal morphology and retrograde transport from endosomes to the Golgi apparatus. Mol. Biol. Cell 24, 2570–2581 (2013).

  22. 22.

    et al. ARF1 and ARF3 are required for the integrity of recycling endosomes and the recycling pathway. Cell Struct. Funct. 37, 141–154 (2012).

  23. 23.

    et al. GBF1: a novel Golgi-associated BFA-resistant guanine nucleotide exchange factor that displays specificity for ADP-ribosylation factor 5. J. Cell Biol. 146, 71–84 (1999).

  24. 24.

    , & ADP-Ribosylation factor 1 (ARF1) regulates recruitment of the AP-3 adaptor complex to membranes. J. Cell Biol. 142, 391–402 (1998).

  25. 25.

    , , , & Overexpression of an ADP-ribosylation factor-guanine nucleotide exchange factor, BIG2, uncouples brefeldin A-induced adaptor protein-1 coat dissociation and membrane tubulation. J. Biol. Chem. 277, 9468–9473 (2002).

  26. 26.

    , , & COPI recruitment ismodulated by a Rab1b-dependent mechanism. Mol. Biol. Cell 14, 2116–2127 (2003).

  27. 27.

    , , , & An activating mutation in ARF1 stabilizes coatomer binding to Golgi membranes. J. Biol. Chem. 269, 3135–3138 (1994).

  28. 28.

    & Activation of ARF6 by ARNO stimulates epithelial cell migration through downstream activation of both Rac1 and phospholipase D. J. Cell Biol. 154, 599–610 (2001).

  29. 29.

    et al. Arf3 is activated uniquely at the trans-Golgi network by brefeldin A-inhibited guanine nucleotide exchange factors. Mol. Biol. Cell 21, 1836–1849 (2010).

  30. 30.

    , & Large Arf1 guanine nucleotide exchange factors: evolution, domain structure, and roles in membrane trafficking and human disease. Mol. Genet. Genom. 282, 329–350 (2009).

  31. 31.

    et al. A Golgi fragmentation pathway in neurodegeneration. Neurobiol. Dis. 29, 221–231 (2008).

  32. 32.

    & Potential role for protein kinases in regulation of bidirectional endoplasmic reticulum-to-Golgi transport revealed by protein kinase inhibitor H89. Mol. Biol. Cell 11, 2577–2590 (2000).

  33. 33.

    , , & Forskolin stimulates detoxification of brefeldin A. J. Biol. Chem. 271, 15870–15873 (1996).

  34. 34.

    , & Regulation of ADP-ribosylation factor 4 expression by small leucine zipper protein and involvement in breast cancer cell migration. Cancer Lett. 314, 185–197 (2012).

  35. 35.

    , , , & The signalling from endoplasmic reticulum-resident bZIP transcription factors involved in diverse cellular physiology. J. Biochem. 149, 507–518 (2011).

  36. 36.

    et al. Activation of OASIS family, ER stress transducers, is dependent on its stabilization. Cell Death Differ. 19, 1939–1949 (2012).

  37. 37.

    et al. JAB1/CSN5 inhibits the activity of Luman/CREB3 by promoting its degradation. Biochim. Biophys. Acta 1829, 921–929 (2013).

  38. 38.

    et al. Chlamydia causes fragmentation of the Golgi compartment to ensure reproduction. Nature 457, 731–735 (2009).

  39. 39.

    et al. Proteolytic elimination of N-myristoyl modifications by the Shigella virulence factor IpaJ. Nature 496, 106–109 (2013).

  40. 40.

    , & Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion. Proc. Natl Acad. Sci. USA 92, 4877–4881 (1995).

  41. 41.

    et al. Chlamydia trachomatis co-opts GBF1 and CERT to acquire host sphingomyelin for distinct roles during intracellular development. PLoS Pathog. 7, e1002198 (2011).

  42. 42.

    et al. A loss-of-function screen reveals Ras- and Raf-independent MEK-ERK signaling during Chlamydia trachomatis infection. Sci. Signal. 3, ra21 (2010).

  43. 43.

    et al. Structurally distinct bacterial TBC-like GAPs link Arf GTPase to Rab1 inactivation to counteract host defenses. Cell 150, 1029–1041 (2012).

  44. 44.

    & Immunology of Chlamydia infection:implications for a Chlamydia trachomatis vaccine. Nat. Rev. Immunol. 5, 149–161 (2005).

  45. 45.

    et al. Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull. World Health Organ. 77, 651–666 (1999).

  46. 46.

    et al. Luman, the cellular counterpart of herpes simplex virus VP16, is processed by regulated intramembrane proteolysis. Mol. Cell Biol. 22, 5639–5649 (2002).

  47. 47.

    et al. Luman is capable of binding and activating transcription from the unfolded protein response element. Biochem. Biophys. Res. Commun. 331, 113–119 (2005).

  48. 48.

    et al. Luman/CREB3 induces transcription of the endoplasmic reticulum (ER) stress response protein Herp through an ER stress response element. Mol. Cell Biol. 26, 7999–8010 (2006).

  49. 49.

    , & The luminal domain of ATF6 senses endoplasmic reticulum (ER) stress and causes translocation of ATF6 from the ER to the Golgi. J. Biol. Chem. 277, 13045–13052 (2002).

  50. 50.

    , , & Role of disulfide bridges formed in the luminal domain of ATF6 in sensing endoplasmic reticulum stress. Mol. Cell Biol. 27, 1027–1043 (2007).

  51. 51.

    et al. Endoplasmic reticulum stress activates cleavage of CREBH to induce a systemic inflammatory response. Cell 124, 587–599 (2006).

  52. 52.

    et al. Unfolded protein response and cell death after depletion of brefeldin A-inhibited guanine nucleotide-exchange protein GBF1. Proc. Natl Acad. Sci. USA 105, 2877–2882 (2008).

  53. 53.

    et al. Golgicide A reveals essential roles for GBF1 in Golgi assembly and function. Nat. Chem. Biol. 5, 157–165 (2009).

  54. 54.

    , , , & Characterization of class I and II ADP-ribosylation factors (Arfs) in live cells: GDP-bound class II Arfs associate with the ER–Golgi intermediate compartment independently of GBF1. Mol. Biol. Cell 19, 3488–3500 (2008).

  55. 55.

    et al. Exo1: a new chemical inhibitor of the exocytic pathway. Proc. Natl Acad. Sci. USA 100, 6469–6474 (2003).

  56. 56.

    , , & Uncoupling of brefeldin a-mediated coatomer protein complex-I dissociation from Golgi redistribution. Traffic 6, 794–802 (2005).

  57. 57.

    & Golgi-disturbing agents. Histochem. Cell Biol. 109, 571–590 (1998).

  58. 58.

    , & Effect of monensin on plant Golgi: re-examination of the monensin-induced changes in cisternal architecture and functional activities of the Golgi apparatus of sycamore suspension-cultured cells. J. Cell Sci. 104, 819–831 (1993).

  59. 59.

    & Golgi structure in stress sensing and apoptosis. Biochim. Biophys. Acta 1744, 406–414 (2005).

  60. 60.

    et al. Novel cis-acting element GASE regulates transcriptional induction by the Golgi stress response. Cell Struct. Funct. 36, 1–12 (2011).

  61. 61.

    , , , & Expansion of the polyQ repeat in ataxin-2 alters its Golgi localization, disrupts the Golgi complex and causes cell death. Human Mol. Genet. 12, 1485–1496 (2003).

  62. 62.

    et al. α-Synuclein impairs macroautophagy: implications for Parkinson’s disease. J. Cell Biol. 190, 1023–1037 (2010).

  63. 63.

    et al. α-synuclein blocks ER–Golgi traffic and Rab1 rescues neuron loss in Parkinson’s models. Science 313, 324–328 (2006).

  64. 64.

    & Consequences of NPC1 and NPC2 loss of function in mammalian neurons. Biochim. Biophys. Acta 1685, 48–62 (2004).

  65. 65.

    , & Fragmentation of the Golgiapparatus in neurodegenerative diseases and cell death. J. Neurol. Sci. 246, 21–30 (2006).

  66. 66.

    , , & IFNgamma inhibits the cytosolic replication of Shigella flexneri via the cytoplasmic RNA sensor RIG-I. PLoS Pathog. 8, e1002809 (2012).

  67. 67.

    , , & CD4+ T cells are necessary and sufficient to confer protection against Chlamydia trachomatis infection in the murine upper genital tract. J. Immunol. 189, 2441–2449 (2012).

  68. 68.

    , & Down-modulation of TCR expression by Salmonella enterica serovar Typhimurium. J. Immunol. 180, 5569–5574 (2008).

  69. 69.

    et al. Compensatory T cell responses in IRG-deficient mice prevent sustained Chlamydia trachomatis infections. PLoS Pathog. 7, e1001346 (2011).

  70. 70.

    & Curr. Protoc. Cell Biol. 48, 11–17 (2010).

Download references

Acknowledgements

We thank J. G. Donaldson for providing the pGEX–VHS–GAT construct, R. A. Weinberg for the Calu-1 cell line and D. Kim for help with confocal microscopy. This work was supported by grants from the US National Institutes of Health (NIH; CA103866) and the D. H. Koch Institute for Integrative Cancer Research to D.M.S. D.M.S. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

    • Jan H. Reiling
    • , Jan E. Carette
    •  & Thijn R. Brummelkamp

    Present addresses: BioMed X GmbH, Im Neuenheimer Feld 583, 69120 Heidelberg, Germany (J.H.R.); Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305, USA (J.E.C.); Department of Biochemistry, Netherlands Cancer Institute, Plesmanlaan 121 1066 CX, Amsterdam, The Netherlands (T.R.B.)

Affiliations

  1. Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA

    • Jan H. Reiling
    • , Sumana Sanyal
    • , Jan E. Carette
    • , Thijn R. Brummelkamp
    • , Hidde L. Ploegh
    •  & David M. Sabatini
  2. Massachusetts Institute of Technology (MIT), Department of Biology, Cambridge, Massachusetts 02142, USA

    • Jan H. Reiling
    • , Sumana Sanyal
    • , Hidde L. Ploegh
    •  & David M. Sabatini
  3. Koch Institute for Integrative Cancer Research, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Jan H. Reiling
    •  & David M. Sabatini
  4. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Andrew J. Olive
    •  & Michael N. Starnbach
  5. Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA

    • David M. Sabatini

Authors

  1. Search for Jan H. Reiling in:

  2. Search for Andrew J. Olive in:

  3. Search for Sumana Sanyal in:

  4. Search for Jan E. Carette in:

  5. Search for Thijn R. Brummelkamp in:

  6. Search for Hidde L. Ploegh in:

  7. Search for Michael N. Starnbach in:

  8. Search for David M. Sabatini in:

Contributions

J.H.R. designed and carried out most of the experiments, analysed data, and wrote the manuscript with input and contributions from all other co-authors. D.M.S. supervised the project, analysed data and edited the manuscript. A.J.O. performed all Chlamydia and Shigella infection assays, edited the manuscript and was supervised by M.N.S. S.S. carried out the pulse-chase labelling and influenza A virus experiments and was supervised by H.L.P. J.E.C. and T.R.B. assisted with haploid genetic screening.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Jan H. Reiling or David M. Sabatini.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb2865

Further reading