Aletrari, Mina-Olga , McKibbin, Craig, Williams, Helen, Pawar, Vidya, Pietroni, Paola, Lord, Mike (J. Mike), Flitsch, Sabine L., Whitehead, Roger C., Swanton, Eileithyia, High, Stephen and Spooner, Robert A. . (2011) Eeyarestatin 1 interferes with both retrograde and anterograde intracellular trafficking pathways. PLoS One, Vol.6 (No.7). e22713. ISSN 1932-6203

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Abstract

Background: The small molecule Eeyarestatin I (ESI) inhibits the endoplasmic reticulum (ER)-cytosol dislocation and subsequent degradation of ERAD (ER associated protein degradation) substrates. Toxins such as ricin and Shiga/Shiga-like toxins (SLTx) are endocytosed and trafficked to the ER. Their catalytic subunits are thought to utilise ERAD-like mechanisms to dislocate from the ER into the cytosol, where a proportion uncouples from the ERAD process, recovers a catalytic conformation and destroys their cellular targets. We therefore investigated ESI as a potential inhibitor of toxin dislocation. Methodology and Principal Findings: Using cytotoxicity measurements, we found no role for ESI as an inhibitor of toxin dislocation from the ER, but instead found that for SLTx, ESI treatment of cells was protective by reducing the rate of toxin delivery to the ER. Microscopy of the trafficking of labelled SLTx and its B chain (lacking the toxic A chain) showed a delay in its accumulation at a peri-nuclear location, confirmed to be the Golgi by examination of SLTx B chain metabolically labelled in the trans-Golgi cisternae. The drug also reduced the rate of endosomal trafficking of diphtheria toxin, which enters the cytosol from acidified endosomes, and delayed the Golgi-specific glycan modifications and eventual plasma membrane appearance of tsO45 VSV-G protein, a classical marker for anterograde trafficking. Conclusions and Significance: ESI acts on one or more components that function during vesicular transport, whilst at least one retrograde trafficking pathway, that of ricin, remains unperturbed.

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Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology
Divisions:	Faculty of Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH):	Ricin, Endoplasmic reticulum, Biological transport
Journal or Publication Title:	PLoS One
Publisher:	Public Library of Science
ISSN:	1932-6203
Date:	25 July 2011
Volume:	Vol.6
Number:	No.7
Page Range:	e22713
Identification Number:	10.1371/journal.pone.0022713
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), National Institutes of Health (U.S.) (NIH), Wellcome Trust (London, England)
Grant number:	BB/D005752/1 (BBSRC), 5U01AI65869-02 (NIH), 080566/Z/06/Z (WT)
References:	1. Spooner RA, Smith DC, Easton AJ, Roberts LM, Lord JM (2006) Retrograde transport pathways utilised by viruses and protein toxins. Virol J 3: 26. 2. Bellisola G, Fracasso G, Ippoliti R, Menestrina G, Rosen A, et al. (2004) Reductive activation of ricin and ricin A-chain immunotoxins by protein disulfide isomerase and thioredoxin reductase. Biochem Pharmacol 67: 1721–1731. 3. Spooner RA, Watson PD, Marsden CJ, Smith DC, Moore KA, et al. (2004) Protein disulphide-isomerase reduces ricin to its A and B chains in the endoplasmic reticulum. Biochem J 383: 285–293. 4. Mayerhofer PU, Cook JP, Wahlman J, Pinheiro TT, Moore KA, et al. (2009) Ricin A chain insertion into endoplasmic reticulum membranes is triggered by a temperature increase to 37uC. J Biol Chem 284: 10232–10242. 5. Ye Y, Shibata Y, Yun C, Ron D, Rapoport TA (2004) A membrane protein complex mediates retro-translocation from the ER lumen into the cytosol. Nature 429: 841–847. 6. Sun F, Zhang R, Gong X, Geng X, Drain PF, et al. (2006) Derlin-1 promotes the efficient degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) and CFTR folding mutants. J Biol Chem 281: 36856–36863. 7. Tsai YC, Weissman AM (2010) The Unfolded Protein Response, Degradation from Endoplasmic Reticulum and Cancer. Genes Cancer 1: 764–778. 8. Rabinovich E, Kerem A, Frohlich KU, Diamant N, Bar-Nun S (2002) AAAATPase p97/Cdc48p, a cytosolic chaperone required for endoplasmic reticulum-associated protein degradation. Mol Cell Biol 22: 626–634. 9. Elkabetz Y, Shapira I, Rabinovich E, Bar-Nun S (2004) Distinct steps in dislocation of luminal endoplasmic reticulum-associated degradation substrates: roles of endoplamic reticulum-bound p97/Cdc48p and proteasome. J Biol Chem 279: 3980–3989. 10. Ye Y, Meyer HH, Rapoport TA (2001) The AAA ATPase Cdc48/p97 and its partners transport proteins from the ER into the cytosol. Nature 414: 652–656. 11. Stirling CJ, Lord JM (2006) Quality control: linking retrotranslocation and degradation. Curr Biol 16: R1035–1037. 12. Wang Q, Li L, Ye Y (2006) Regulation of retrotranslocation by p97-associated deubiquitinating enzyme ataxin-3. J Cell Biol 174: 963–971. 13. Zhong X, Pittman RN (2006) Ataxin-3 binds VCP/p97 and regulates retrotranslocation of ERAD substrates. Hum Mol Genet 15: 2409–2420. 14. Medicherla B, Kostova Z, Schaefer A, Wolf DH (2004) A genomic screen identifies Dsk2p and Rad23p as essential components of ER-associated degradation. EMBO Rep 5: 692–697. 15. Wesche J, Rapak A, Olsnes S (1999) Dependence of ricin toxicity on translocation of the toxin A-chain from the endoplasmic reticulum to the cytosol. J Biol Chem 274: 34443–34449. 16. Willer M, Forte GM, Stirling CJ (2008) Sec61p is required for ERAD-L: genetic dissection of the translocation and ERAD-L functions of Sec61P using novel derivatives of CPY. J Biol Chem 283: 33883–33888. 17. Slominska-Wojewodzka M, Gregers TF, Walchli S, Sandvig K (2006) EDEM is involved in retrotranslocation of ricin from the endoplasmic reticulum to the cytosol. Mol Biol Cell 17: 1664–1675. 18. Redmann V, Oresic K, Tortorella LL, Cook JP, Lord M, et al. (2011) Dislocation of ricin toxin A chains in human cells utilizes selective cellular factors. J Biol Chem 286: 21231–21238. 19. Spooner RA, Hart PJ, Cook JP, Pietroni P, Rogon C, et al. (2008) Cytosolic chaperones influence the fate of a toxin dislocated from the endoplasmic reticulum. Proc Natl Acad Sci U S A 105: 17408–17413. 20. Endo Y, Tsurugi K (1987) RNA N-glycosidase activity of ricin A-chain. Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. J Biol Chem 262: 8128–8130. 21. Li S, Spooner RA, Allen SC, Guise CP, Ladds G, et al. (2010) Foldingcompetent and folding-defective forms of ricin A chain have different fates after retrotranslocation from the endoplasmic reticulum. Mol Biol Cell 21: 2543–2554. 22. Endo Y, Tsurugi K, Yutsudo T, Takeda Y, Ogasawara T, et al. (1988) Site of action of a Vero toxin (VT2) from Escherichia coli O157:H7 and of Shiga toxin on eukaryotic ribosomes. RNA N-glycosidase activity of the toxins. Eur J Biochem 171: 45–50. 23. Yu M, Haslam DB (2005) Shiga toxin is transported from the endoplasmic reticulum following interaction with the luminal chaperone HEDJ/ERdj3. Infect Immun 73: 2524–2532. 24. Falguieres T, Johannes L (2006) Shiga toxin B-subunit binds to the chaperone BiP and the nucleolar protein B23. Biol Cell 98: 125–134. 25. Williams DB, Watts TH (1995) Molecular chaperones in antigen presentation. Curr Opin Immunol 7: 77–84. 26. Cresswell P (2000) Intracellular surveillance: controlling the assembly of MHC class I-peptide complexes. Traffic 1: 301–305. 27. Tortorella D, Gewurz BE, Furman MH, Schust DJ, Ploegh HL (2000) Viral subversion of the immune system. Annu Rev Immunol 18: 861–926. 28. Wiertz EJ, Jones TR, Sun L, Bogyo M, Geuze HJ, et al. (1996) The human cytomegalovirus US11 gene product dislocates MHC class I heavy chains from the endoplasmic reticulum to the cytosol. Cell 84: 769–779. 29. Mueller B, Lilley BN, Ploegh HL (2006) SEL1L, the homologue of yeast Hrd3p, is involved in protein dislocation from the mammalian ER. J Cell Biol 175: 261–270. 30. Mueller B, Klemm EJ, Spooner E, Claessen JH, Ploegh HL (2008) SEL1L nucleates a protein complex required for dislocation of misfolded glycoproteins. Proc Natl Acad Sci U S A 105: 12325–12330. 31. Baker BM, Tortorella D (2007) Dislocation of an endoplasmic reticulum membrane glycoprotein involves the formation of partially dislocated ubiquitinated polypeptides. J Biol Chem 282: 26845–26856. 32. Schulze A, Standera S, Buerger E, Kikkert M, van Voorden S, et al. (2005) The ubiquitin-domain protein HERP forms a complex with components of the endoplasmic reticulum associated degradation pathway. J Mol Biol 354: 1021–1027. 33. Burr ML, Cano F, Svobodova S, Boyle LH, Boname JM, et al. (2011) HRD1 and UBE2J1 target misfolded MHC class I heavy chains for endoplasmic reticulum-associated degradation. Proc Natl Acad Sci U S A 108: 2034–2039. 34. Fiebiger E, Hirsch C, Vyas JM, Gordon E, Ploegh HL, et al. (2004) Dissection of the dislocation pathway for type I membrane proteins with a new small molecule inhibitor, eeyarestatin. Mol Biol Cell 15: 1635–1646. 35. Wang Q, Li L, Ye Y (2008) Inhibition of p97-dependent protein degradation by Eeyarestatin I. J Biol Chem 283: 7445–7454. 36. Wang Q, Shinkre BA, Lee JG, Weniger MA, Liu Y, et al. (2010) The ERAD inhibitor Eeyarestatin I is a bifunctional compound with a membrane-binding domain and a p97/VCP inhibitory group. PLoS One 5: e15479. 37. Dolan BP, Li L, Veltri CA, Ireland CM, Bennink JR, et al. (2011) Distinct pathways generate peptides from defective ribosomal products for CD8+ T cell immunosurveillance. J Immunol 186: 2065–2072. 38. Cross BC, McKibbin C, Callan AC, Roboti P, Piacenti M, et al. (2009) Eeyarestatin I inhibits Sec61-mediated protein translocation at the endoplasmic reticulum. J Cell Sci 122: 4393–4400. 39. Ernst R, Claessen JHL, Mueller B, Sanyal S, Spooner E, et al. (2011) Enzymatic blockade of the ubiquitin-proteasome pathway. PLoS Biol 8: e10000605. 40. Craiu A, Gaczynska M, Akopian T, Gramm CF, Fenteany G, et al. (1997) Lactacystin and clasto-lactacystin beta-lactone modify multiple proteasome betasubunits and inhibit intracellular protein degradation and major histocompatibility complex class I antigen presentation. J Biol Chem 272: 13437–13445. 41. Tam PJ, Lingwood CA (2007) Membrane cytosolic translocation of verotoxin A1 subunit in target cells. Microbiology 153: 2700–2710. 42. Hudson TH, Neville DM, Jr. (1987) Temporal separation of protein toxin translocation from processing events. J Biol Chem 262: 16484–16494. 43. Neville DM, Jr., Youle RJ (1982) Monoclonal antibody-ricin or ricin A chain hybrids: kinetic analysis of cell killing for tumor therapy. Immunol Rev 62: 75–91. 44. Sandvig K, Ryd M, Garred O, Schweda E, Holm PK, et al. (1994) Retrograde transport from the Golgi complex to the ER of both Shiga toxin and the nontoxic Shiga B-fragment is regulated by butyric acid and cAMP. J Cell Biol 126: 53–64. 45. Sandvig K, Garred O, Prydz K, Kozlov JV, Hansen SH, et al. (1992) Retrograde transport of endocytosed Shiga toxin to the endoplasmic reticulum. Nature 358: 510–512. 46. Mallard F, Antony C, Tenza D, Salamero J, Goud B, et al. (1998) Direct pathway from early/recycling endosomes to the Golgi apparatus revealed through the study of shiga toxin B-fragment transport. J Cell Biol 143: 973–990. 47. Spooner RA, Watson P, Smith DC, Boal F, Amessou M, et al. (2008) The secretion inhibitor Exo2 perturbs trafficking of Shiga toxin between endosomes and the trans-Golgi network. Biochem J 414: 471–484. 48. Blewitt MG, Chung LA, London E (1985) Effect of pH on the conformation of diphtheria toxin and its implications for membrane penetration. Biochemistry 24: 5458–5464. 49. Gallione CJ, Rose JK (1985) A single amino acid substitution in a hydrophobic domain causes temperature-sensitive cell-surface transport of a mutant viral glycoprotein. J Virol 54: 374–382. 50. Presley JF, Cole NB, Schroer TA, Hirschberg K, Zaal KJ, et al. (1997) ER-to- Golgi transport visualized in living cells. Nature 389: 81–85. 51. Hammond C, Helenius A (1994) Quality control in the secretory pathway: retention of a misfolded viral membrane glycoprotein involves cycling between the ER, intermediate compartment, and Golgi apparatus. J Cell Biol 126: 41–52. 52. Donaldson JG, Lippincott-Schwartz J, Bloom GS, Kreis TE, Klausner RD (1990) Dissociation of a 110-kD peripheral membrane protein from the Golgi apparatus is an early event in brefeldin A action. J Cell Biol 111: 2295–2306. 53. Orci L, Tagaya M, Amherdt M, Perrelet A, Donaldson JG, et al. (1991) Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrincoated buds on Golgi cisternae. Cell 64: 1183–1195. 54. Lippincott-Schwartz J, Yuan LC, Bonifacino JS, Klausner RD (1989) Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell 56: 801–813. 55. Chen SS, Huang AS (1986) Further characterization of the vesicular stomatitis virus temperature-sensitive O45 mutant: intracellular conversion of the glycoprotein to a soluble form. J Virol 59: 210–215. 56. Scheel J, Pepperkok R, Lowe M, Griffiths G, Kreis TE (1997) Dissociation of coatomer from membranes is required for brefeldin A-induced transfer of Golgi enzymes to the endoplasmic reticulum. J Cell Biol 137: 319–333. 57. Wang Q, Mora-Jensen H, Weniger MA, Perez-Galan P, Wolford C, et al. (2009) ERAD inhibitors integrate ER stress with an epigenetic mechanism to activate BH3-only protein NOXA in cancer cells. Proc Natl Acad Sci U S A 106: 2200–2205. 58. Kar R, Singha PK, Venkatachalam MA, Saikumar P (2009) A novel role for MAP1 LC3 in nonautophagic cytoplasmic vacuolation death of cancer cells. Oncogene 28: 2556–2568. 59. White J, Johannes L, Mallard F, Girod A, Grill S, et al. (1999) Rab6 coordinates a novel Golgi to ER retrograde transport pathway in live cells. J Cell Biol 147: 743–760. 60. Monier S, Jollivet F, Janoueix-Lerosey I, Johannes L, Goud B (2002) Characterization of novel Rab6-interacting proteins involved in endosome-to- TGN transport. Traffic 3: 289–297. 61. Del Nery E, Miserey-Lenkei S, Falguieres T, Nizak C, Johannes L, et al. (2006) Rab6A and Rab6A’ GTPases play non-overlapping roles in membrane trafficking. Traffic 7: 394–407. 62. Chen A, AbuJarour RJ, Draper RK (2003) Evidence that the transport of ricin to the cytoplasm is independent of both Rab6A and COPI. J Cell Sci 116: 3503–3510. 63. Amessou M, Fradagrada A, Falguieres T, Lord JM, Smith DC, et al. (2007) Syntaxin 16 and syntaxin 5 are required for efficient retrograde transport of several exogenous and endogenous cargo proteins. J Cell Sci 120: 1457–1468. 64. Sko¨nland SS, Walchli S, Utskarpen A, Wandinger-Ness A, Sandvig K (2007) Phosphoinositide-regulated retrograde transport of ricin: crosstalk between hVps34 and sorting nexins. Traffic 8: 297–309. 65. Dyve AB, Bergan J, Utskarpen A, Sandvig K (2009) Sorting nexin 8 regulates endosome-to-Golgi transport. Biochem Biophys Res Commun 390: 109–114. 66. Bujny MV, Popoff V, Johannes L, Cullen PJ (2007) The retromer component sorting nexin-1 is required for efficient retrograde transport of Shiga toxin from early endosome to the trans Golgi network. J Cell Sci 120: 2010–2021. 67. Popoff V, Mardones GA, Bai SK, Chambon V, Tenza D, et al. (2009) Analysis of articulation between clathrin and retromer in retrograde sorting on early endosomes. Traffic 10: 1868–1880. 68. Smith DC, Spooner RA, Watson PD, Murray JL, Hodge TW, et al. (2006) Internalized Pseudomonas exotoxin A can exploit multiple pathways to reach the endoplasmic reticulum. Traffic 7: 379–393. 69. Chadfield MS, Hinton MH (2004) In vitro activity of nitrofuran derivatives on growth and morphology of Salmonella enterica serotype Enteritidis. J Appl Microbiol 96: 1002–1012. 70. Polycarpou-Schwarz M, Muller K, Denger S, Riddell A, Lewis J, et al. (2007) Thanatop: a novel 5-nitrofuran that is a highly active, cell-permeable inhibitor of topoisomerase II. Cancer Res 67: 4451–4458. 71. Maya JD, Bollo S, Nunez-Vergara LJ, Squella JA, Repetto Y, et al. (2003) Trypanosoma cruzi: effect and mode of action of nitroimidazole and nitrofuran derivatives. Biochem Pharmacol 65: 999–1006. 72. Letelier ME, Izquierdo P, Godoy L, Lepe AM, Faundez M (2004) Liver microsomal biotransformation of nitro-aryl drugs: mechanism for potential oxidative stress induction. J Appl Toxicol 24: 519–525. 73. Lakkaraju AK, Luyet PP, Parone P, Falguieres T, Strub K (2007) Inefficient targeting to the endoplasmic reticulum by the signal recognition particle elicits selective defects in post-ER membrane trafficking. Exp Cell Res 313: 834–847. 74. Clague MJ, Urbe S (2006) Endocytosis: the DUB version. Trends Cell Biol 16: 551–559. 75. Wakana Y, Takai S, Nakajima K, Tani K, Yamamoto A, et al. (2008) Bap31 is an itinerant protein that moves between the peripheral endoplasmic reticulum (ER) and a juxtanuclear compartment related to ER-associated Degradation. Mol Biol Cell 19: 1825–1836. 76. Hosokawa N, You Z, Tremblay LO, Nagata K, Herscovics A (2007) Stimulation of ERAD of misfolded null Hong Kong alpha1-antitrypsin by Golgi alpha1,2- mannosidases. Biochem Biophys Res Commun 362: 626–632. 77. Vashist S, Kim W, Belden WJ, Spear ED, Barlowe C, et al. (2001) Distinct retrieval and retention mechanisms are required for the quality control of endoplasmic reticulum protein folding. J Cell Biol 155: 355–368. 78. Fu L, Sztul E (2003) Traffic-independent function of the Sar1p/COPII machinery in proteasomal sorting of the cystic fibrosis transmembrane conductance regulator. J Cell Biol 160: 157–163. 79. Aghi M, Kramm CM, Chou TC, Breakefield XO, Chiocca EA (1998) Synergistic anticancer effects of ganciclovir/thymidine kinase and 5-fluorocytosine/ cytosine deaminase gene therapies. J Natl Cancer Inst 90: 370–380. 80. Chiu CF, Ghanekar Y, Frost L, Diao A, Morrison D, et al. (2008) ZFPL1, a novel ring finger protein required for cis-Golgi integrity and efficient ER-to- Golgi transport. Embo J 27: 934–947. 81. Schagger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368–379.
URI:	http://wrap.warwick.ac.uk/id/eprint/38354

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