Reprogramming the unfolded protein response for replication by porcine reproductive and respiratory syndrome virus


Autoři: Peng Gao aff001;  Yue Chai aff001;  Jiangwei Song aff001;  Teng Liu aff001;  Peng Chen aff001;  Lei Zhou aff001;  Xinna Ge aff001;  Xin Guo aff001;  Jun Han aff001;  Hanchun Yang aff001
Působiště autorů: Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, China Agricultural University College of Veterinary Medicine, Beijing, People’s Republic of China aff001
Vyšlo v časopise: Reprogramming the unfolded protein response for replication by porcine reproductive and respiratory syndrome virus. PLoS Pathog 15(11): e32767. doi:10.1371/journal.ppat.1008169
Kategorie: Research Article
doi: 10.1371/journal.ppat.1008169

Souhrn

The unfolded protein response (UPR) in the endoplasmic reticulum (ER) constitutes a critical component of host innate immunity against microbial infections. In this report, we show that porcine reproductive and respiratory syndrome virus (PRRSV) utilizes the UPR machinery for its own benefit. We provide evidence that the virus targets the UPR central regulator GRP78 for proteasomal degradation via a mechanism that requires viral glycoprotein GP2a, while both IRE1-XBP1s and PERK-eIF2α-ATF4 signaling branches of the UPR are turned on at early stage of infection. The activated effector XBP1s was found to enter the nucleus, but ATF4 was unexpectedly diverted to cytoplasmic viral replication complexes by means of nonstructural proteins nsp2/3 to promote viral RNA synthesis. RNAi knockdown of either ATF4 or XBP1s dramatically attenuated virus titers, while overexpression caused increases. These observations reveal attractive host targets (e.g., ATF4 and XBP1s) for antiviral drugs and have implications in vaccine development.

Klíčová slova:

Antibodies – Confocal microscopy – Messenger RNA – RNA synthesis – RNA viruses – Small interfering RNAs – Transfection – Viral replication


Zdroje

1. Walter P, Ron D. The unfolded protein response: from stress pathway to homeostatic regulation. Science. 2011;334:1081–1086. doi: 10.1126/science.1209038 22116877

2. Schroder M. The unfolded protein response. Mol Biotechnol. 2006;34:279–290. doi: 10.1385/MB:34:2:279 17172673

3. Bernales S, Papa FR, Walter P. Intracellular signaling by the unfolded protein response. Annu Rev Cell Dev Biol. 2006;22:487–508. doi: 10.1146/annurev.cellbio.21.122303.120200 16822172

4. Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol. 2007;8:519–529. doi: 10.1038/nrm2199 17565364

5. Hetz C, Papa FR. The Unfolded Protein Response and Cell Fate Control. Mol Cell. 2018;69:169–181. doi: 10.1016/j.molcel.2017.06.017 29107536

6. Bertolotti A, Zhang Y, Hendershot LM, Harding HP, Ron D. Dynamic interaction of BiP and ER stress transducers in the unfolded-protein response. Nat Cell Biol. 2000;2:326–332. doi: 10.1038/35014014 10854322

7. Lewy TG, Grabowski JM, Bloom ME. BiP: Master Regulator of the Unfolded Protein Response and Crucial Factor in Flavivirus Biology. Yale J Biol Med. 2017;90:291–300. 28656015

8. Okamura K, Kimata Y, Higashio H, Tsuru A, Kohno K. Dissociation of Kar2p/BiP from an ER sensory molecule, Ire1p, triggers the unfolded protein response in yeast. Biochem Biophys Res Commun. 2000;279:445–450. doi: 10.1006/bbrc.2000.3987 11118306

9. Wek RC, Jiang HY, Anthony TG. Coping with stress: eIF2 kinases and translational control. Biochem Soc Trans. 2006;34:7–11. doi: 10.1042/BST20060007 16246168

10. Vattem KM, Wek RC. Reinitiation involving upstream ORFs regulates ATF4 mRNA translation in mammalian cells. Proc Natl Acad Sci U S A. 2004;101:11269–11274. doi: 10.1073/pnas.0400541101 15277680

11. Harding HP, Novoa I, Zhang Y, Zeng H, Wek R, Schapira M, et al. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell. 2000;6:1099–1108. doi: 10.1016/s1097-2765(00)00108-8 11106749

12. Harding HP, Zhang Y, Zeng H, Novoa I, Lu PD, Calfon M, et al. An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol Cell. 2003;11:619–633. doi: 10.1016/s1097-2765(03)00105-9 12667446

13. Fusakio ME, Willy JA, Wang Y, Mirek ET, Al Baghdadi RJ, Adams CM, et al. Transcription factor ATF4 directs basal and stress-induced gene expression in the unfolded protein response and cholesterol metabolism in the liver. Mol Biol Cell. 2016;27:1536–1551. doi: 10.1091/mbc.E16-01-0039 26960794

14. Ohoka N, Yoshii S, Hattori T, Onozaki K, Hayashi H. TRB3, a novel ER stress-inducible gene, is induced via ATF4-CHOP pathway and is involved in cell death. EMBO J. 2005;24:1243–1255. doi: 10.1038/sj.emboj.7600596 15775988

15. Ameri K, Harris AL. Activating transcription factor 4. Int J Biochem Cell Biol. 2008;40:14–21. doi: 10.1016/j.biocel.2007.01.020 17466566

16. Schindler AJ, Schekman R. In vitro reconstitution of ER-stress induced ATF6 transport in COPII vesicles. Proc Natl Acad Sci U S A. 2009;106:17775–17780. doi: 10.1073/pnas.0910342106 19822759

17. Chen X, Shen J, Prywes R. The luminal domain of ATF6 senses endoplasmic reticulum (ER) stress and causes translocation of ATF6 from the ER to the Golgi. J Biol Chem. 2002;277:13045–13052. doi: 10.1074/jbc.M110636200 11821395

18. Hong M, Luo S, Baumeister P, Huang JM, Gogia RK, Li M, et al. Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. J Biol Chem. 2004;279:11354–11363. doi: 10.1074/jbc.M309804200 14699159

19. Adachi Y, Yamamoto K, Okada T, Yoshida H, Harada A, Mori K. ATF6 Is a Transcription Factor Specializing in the Regulation of Quality Control Proteins in the Endoplasmic Reticulum. Cell Structure And Function. 2008;33:75–89. doi: 10.1247/csf.07044 18360008

20. Back SH, Schroder M, Lee K, Zhang K, Kaufman RJ. ER stress signaling by regulated splicing: IRE1/HAC1/XBP1. Methods. 2005;35:395–416. doi: 10.1016/j.ymeth.2005.03.001 15804613

21. Patil C, Walter P. Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr Opin Cell Biol. 2001;13:349–355. doi: 10.1016/s0955-0674(00)00219-2 11343907

22. Hollien J, Weissman JS. Decay of endoplasmic reticulum-localized mRNAs during the unfolded protein response. Science. 2006;313:104–107. doi: 10.1126/science.1129631 16825573

23. Sakaki K, Yoshina S, Shen X, Han J, DeSantis MR, Xiong M, et al. RNA surveillance is required for endoplasmic reticulum homeostasis. Proc Natl Acad Sci U S A. 2012;109:8079–8084. doi: 10.1073/pnas.1110589109 22562797

24. Sriburi R, Jackowski S, Mori K, Brewer JW. XBP1: a link between the unfolded protein response, lipid biosynthesis, and biogenesis of the endoplasmic reticulum. J Cell Biol. 2004;167:35–41. doi: 10.1083/jcb.200406136 15466483

25. Yang J, Wu X, Wu X, Zhou D, Lin T, Ding S, et al. The Multiple Roles of XBP1 in Regulation of Glucose and Lipid Metabolism. Curr Protein Pept Sci. 2017;18:630–635. doi: 10.2174/1389203717666160627085011 27356931

26. Chakrabarti A, Chen AW, Varner JD. A review of the mammalian unfolded protein response. Biotechnol Bioeng. 2011;108:2777–2793. doi: 10.1002/bit.23282 21809331

27. Frabutt DA, Wang B, Riaz S, Schwartz RC, Zheng YH. Innate Sensing of Influenza A Virus Hemagglutinin Glycoproteins by the Host Endoplasmic Reticulum (ER) Stress Pathway Triggers a Potent Antiviral Response via ER-Associated Protein Degradation. J Virol. 2018;92: e01690–01617. doi: 10.1128/JVI.01690-17 29046440

28. Zhang HM, Dai H, Hanson PJ, Li H, Guo H, Ye X, et al. Antiviral activity of an isatin derivative via induction of PERK-Nrf2-mediated suppression of cap-independent translation. ACS Chem Biol. 2014;9:1015–1024. doi: 10.1021/cb400775z 24547890

29. So JS. Roles of Endoplasmic Reticulum Stress in Immune Responses. Mol Cells. 2018;41:705–716. doi: 10.14348/molcells.2018.0241 30078231

30. Dandekar A, Mendez R, Zhang K. Cross talk between ER stress, oxidative stress, and inflammation in health and disease. Methods Mol Biol. 2015;1292:205–214. doi: 10.1007/978-1-4939-2522-3_15 25804758

31. Song S, Tan J, Miao Y, Zhang Q. Crosstalk of ER stress-mediated autophagy and ER-phagy: Involvement of UPR and the core autophagy machinery. J Cell Physiol. 2018;233:3867–3874. doi: 10.1002/jcp.26137 28777470

32. Han CY, Rho HS, Kim A, Kim TH, Jang K, Jun DW, et al. FXR Inhibits Endoplasmic Reticulum Stress-Induced NLRP3 Inflammasome in Hepatocytes and Ameliorates Liver Injury. Cell Rep. 2018;24:2985–2999. doi: 10.1016/j.celrep.2018.07.068 30208322

33. Li S, Kong L, Yu X. The expanding roles of endoplasmic reticulum stress in virus replication and pathogenesis. Crit Rev Microbiol. 2015;41:150–164. doi: 10.3109/1040841X.2013.813899 25168431

34. Cavanagh D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae. Arch Virol. 1997;142:629–633. 9349308

35. Kuhn JH, Lauck M, Bailey AL, Shchetinin AM, Vishnevskaya TV, Bao Y, et al. Reorganization and expansion of the nidoviral family Arteriviridae. Arch Virol. 2016;161:755–768. doi: 10.1007/s00705-015-2672-z 26608064

36. Wensvoort G, Terpstra C, Pol JM, ter Laak EA, Bloemraad M, de Kluyver EP, et al. Mystery swine disease in The Netherlands: the isolation of Lelystad virus. Vet Q. 1991;13:121–130. doi: 10.1080/01652176.1991.9694296 1835211

37. Han J, Zhou L, Ge X, Guo X, Yang H. Pathogenesis and control of the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus. Vet Microbiol. 2017;209:30–47. doi: 10.1016/j.vetmic.2017.02.020 28292547

38. Tian K, Yu X, Zhao T, Feng Y, Cao Z, Wang C, et al. Emergence of fatal PRRSV variants: unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark. PLoS One. 2007;2:e526. doi: 10.1371/journal.pone.0000526 17565379

39. Zhao K, Ye C, Chang XB, Jiang CG, Wang SJ, Cai XH, et al. Importation and Recombination Are Responsible for the Latest Emergence of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus in China. J Virol. 2015;89:10712–10716. doi: 10.1128/JVI.01446-15 26246582

40. Wills RW, Zimmerman JJ, Yoon KJ, Swenson SL, McGinley MJ, Hill HT, et al. Porcine reproductive and respiratory syndrome virus: a persistent infection. Vet Microbiol. 1997;55:231–240. doi: 10.1016/s0378-1135(96)01337-5 9220618

41. Chen WY, Schniztlein WM, Calzada-Nova G, Zuckermann FA. Genotype 2 Strains of Porcine Reproductive and Respiratory Syndrome Virus Dysregulate Alveolar Macrophage Cytokine Production via the Unfolded Protein Response. J Virol. 2018;92(2): e01251–17. doi: 10.1128/JVI.01251-17 29070690

42. Lee AS. The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress. Methods. 2005;35:373–381. doi: 10.1016/j.ymeth.2004.10.010 15804610

43. Su HL, Liao CL, Lin YL. Japanese encephalitis virus infection initiates endoplasmic reticulum stress and an unfolded protein response. J Virol. 2002;76:4162–4171. doi: 10.1128/JVI.76.9.4162-4171.2002 11932381

44. Jheng JR, Lau KS, Tang WF, Wu MS, Horng JT. Endoplasmic reticulum stress is induced and modulated by enterovirus 71. Cell Microbiol. 2010;12:796–813. doi: 10.1111/j.1462-5822.2010.01434.x 20070307

45. Neerukonda SN, Katneni UK, Bott M, Golovan SP, Parcells MS. Induction of the unfolded protein response (UPR) during Marek's disease virus (MDV) infection. Virology. 2018;522:1–12. doi: 10.1016/j.virol.2018.06.016 29979959

46. Vekich JA, Belmont PJ, Thuerauf DJ, Glembotski CC. Protein disulfide isomerase-associated 6 is an ATF6-inducible ER stress response protein that protects cardiac myocytes from ischemia/reperfusion-mediated cell death. J Mol Cell Cardiol. 2012;53:259–267. doi: 10.1016/j.yjmcc.2012.05.005 22609432

47. Fang Y, Snijder EJ. The PRRSV replicase: exploring the multifunctionality of an intriguing set of nonstructural proteins. Virus Res. 2010;154:61–76. doi: 10.1016/j.virusres.2010.07.030 20696193

48. Kilberg MS, Shan J, Su N. ATF4-dependent transcription mediates signaling of amino acid limitation. Trends Endocrinol Metab. 2009;20:436–443. doi: 10.1016/j.tem.2009.05.008 19800252

49. van der Meer Y, van Tol H, Locker JK, Snijder EJ. ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex. J Virol. 1998;72:6689–6698. 9658116

50. Snijder EJ, van Tol H, Roos N, Pedersen KW. Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex. J Gen Virol. 2001;82:985–994. doi: 10.1099/0022-1317-82-5-985 11297673

51. Huo Y, Fan L, Yin S, Dong Y, Guo X, Yang H, et al. Involvement of unfolded protein response, p53 and Akt in modulation of porcine reproductive and respiratory syndrome virus-mediated JNK activation. Virology. 2013;444:233–240. doi: 10.1016/j.virol.2013.06.015 23850458

52. Qi L, Tsai B, Arvan P. New Insights into the Physiological Role of Endoplasmic Reticulum-Associated Degradation. Trends Cell Biol. 2017;27:430–440. doi: 10.1016/j.tcb.2016.12.002 28131647

53. Jheng JR, Ho JY, Horng JT. ER stress, autophagy, and RNA viruses. Front Microbiol. 2014;5:388. doi: 10.3389/fmicb.2014.00388 25140166

54. Verchot J. How does the stressed out ER find relief during virus infection? Curr Opin Virol. 2016;17:74–79. doi: 10.1016/j.coviro.2016.01.018 26871502

55. Fung TS, Liu DX. Coronavirus infection, ER stress, apoptosis and innate immunity. Front Microbiol. 2014;5:296. doi: 10.3389/fmicb.2014.00296 24987391

56. He B. Viruses, endoplasmic reticulum stress, and interferon responses. Cell Death Differ. 2006;13:393–403. doi: 10.1038/sj.cdd.4401833 16397582

57. Wei D, Li NL, Zeng Y, Liu B, Kumthip K, Wang TT, et al. The Molecular Chaperone GRP78 Contributes to Toll-like Receptor 3-mediated Innate Immune Response to Hepatitis C Virus in Hepatocytes. J Biol Chem. 2016;291:12294–12309. doi: 10.1074/jbc.M115.711598 27129228

58. Zhang L, Li Z, Ding G, La X, Yang P, Li Z. GRP78 plays an integral role in tumor cell inflammation-related migration induced by M2 macrophages. Cell Signal. 2017;37:136–148. doi: 10.1016/j.cellsig.2017.06.008 28629783

59. Medigeshi GR, Lancaster AM, Hirsch AJ, Briese T, Lipkin WI, Defilippis V, et al. West Nile virus infection activates the unfolded protein response, leading to CHOP induction and apoptosis. J Virol. 2007;81:10849–10860. doi: 10.1128/JVI.01151-07 17686866

60. Liao Y, Fung TS, Huang M, Fang SG, Zhong Y, Liu DX. Upregulation of CHOP/GADD153 during coronavirus infectious bronchitis virus infection modulates apoptosis by restricting activation of the extracellular signal-regulated kinase pathway. J Virol. 2013;87:8124–8134. doi: 10.1128/JVI.00626-13 23678184

61. Cheng G, Feng Z, He B. Herpes simplex virus 1 infection activates the endoplasmic reticulum resident kinase PERK and mediates eIF-2alpha dephosphorylation by the gamma(1)34.5 protein. J Virol. 2005;79:1379–1388. doi: 10.1128/JVI.79.3.1379-1388.2005 15650164

62. Perera N, Miller JL, Zitzmann N. The role of the unfolded protein response in dengue virus pathogenesis. Cell Microbiol. 2017;19:e12734.

63. Jiang G, Santos Rocha C, Hirao LA, Mendes EA, Tang Y, Thompson GR 3rd, et al. HIV Exploits Antiviral Host Innate GCN2-ATF4 Signaling for Establishing Viral Replication Early in Infection. MBio. 2017;8: e01518–01516. doi: 10.1128/mBio.01518-16 28465428

64. Bechill J, Chen Z, Brewer JW, Baker SC. Coronavirus infection modulates the unfolded protein response and mediates sustained translational repression. J Virol. 2008;82:4492–4501. doi: 10.1128/JVI.00017-08 18305036

65. Xu L, Khadijah S, Fang S, Wang L, Tay FP, Liu DX. The cellular RNA helicase DDX1 interacts with coronavirus nonstructural protein 14 and enhances viral replication. J Virol. 2010;84:8571–8583. doi: 10.1128/JVI.00392-10 20573827

66. Wu CH, Chen PJ, Yeh SH. Nucleocapsid phosphorylation and RNA helicase DDX1 recruitment enables coronavirus transition from discontinuous to continuous transcription. Cell Host Microbe. 2014;16:462–472. doi: 10.1016/j.chom.2014.09.009 25299332

67. Singleton DC, Harris AL. Targeting the ATF4 pathway in cancer therapy. Expert Opin Ther Targets. 2012;16:1189–1202. doi: 10.1517/14728222.2012.728207 23009153

68. Gao S, Ge A, Xu S, You Z, Ning S, Zhao Y, et al. PSAT1 is regulated by ATF4 and enhances cell proliferation via the GSK3beta/beta-catenin/cyclin D1 signaling pathway in ER-negative breast cancer. J Exp Clin Cancer Res. 2017;36:179. doi: 10.1186/s13046-017-0648-4 29216929

69. Rzymski T, Milani M, Singleton DC, Harris AL. Role of ATF4 in regulation of autophagy and resistance to drugs and hypoxia. Cell Cycle. 2009;8:3838–3847. doi: 10.4161/cc.8.23.10086 19887912

70. Fung TS, Liao Y, Liu DX. The endoplasmic reticulum stress sensor IRE1alpha protects cells from apoptosis induced by the coronavirus infectious bronchitis virus. J Virol. 2014;88:12752–12764. doi: 10.1128/JVI.02138-14 25142592

71. Margariti A, Li H, Chen T, Martin D, Vizcay-Barrena G, Alam S, et al. XBP1 mRNA splicing triggers an autophagic response in endothelial cells through BECLIN-1 transcriptional activation. The Journal of biological chemistry. 2013;288(2):859–72. doi: 10.1074/jbc.M112.412783 23184933

72. Bechill J, Chen Z, Brewer JW, Baker SC. Mouse hepatitis virus infection activates the Ire1/XBP1 pathway of the unfolded protein response. Advances in experimental medicine and biology. 2006;581:139–44. doi: 10.1007/978-0-387-33012-9_24 17037520

73. Sharma M, Bhattacharyya S, Sharma KB, Chauhan S, Asthana S, Abdin MZ, et al. Japanese encephalitis virus activates autophagy through XBP1 and ATF6 ER stress sensors in neuronal cells. J Gen Virol. 2017;98:1027–1039. doi: 10.1099/jgv.0.000792 28535855

74. Friedlander R, Jarosch E, Urban J, Volkwein C, Sommer T. A regulatory link between ER-associated protein degradation and the unfolded-protein response. Nat Cell Biol. 2000;2:379–384. doi: 10.1038/35017001 10878801

75. Xue M, Fu F, Ma Y, Zhang X, Li L, Feng L, et al. The PERK Arm of the Unfolded Protein Response Negatively Regulates Transmissible Gastroenteritis Virus Replication by Suppressing Protein Translation and Promoting Type I Interferon Production. J Virol. 2018;92(15):e00431–18. doi: 10.1128/JVI.00431-18 29769338

76. Zhou L, Wang Z, Ding Y, Ge X, Guo X, Yang H. NADC30-like Strain of Porcine Reproductive and Respiratory Syndrome Virus, China. Emerg Infect Dis. 2015;21:2256–2257. doi: 10.3201/eid2112.150360 26584305

77. Zhou L, Zhang J, Zeng J, Yin S, Li Y, Zheng L, et al. The 30-amino-acid deletion in the Nsp2 of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China is not related to its virulence. J Virol. 2009;83:5156–5167. doi: 10.1128/JVI.02678-08 19244318

78. Li J, Jin Z, Gao Y, Zhou L, Ge X, Guo X, et al. Development of the full-length cDNA clones of two porcine epidemic diarrhea disease virus isolates with different virulence. PLoS One. 2017;12:e0173998. doi: 10.1371/journal.pone.0173998 28301551

79. Zhang GQ, Ge XN, Guo X, Yang HC. Genomic analysis of two porcine encephalomyocarditis virus strains isolated in China. Arch Virol. 2007;152:1209–1213. doi: 10.1007/s00705-006-0930-9 17294091

80. van Dinten LC, den Boon JA, Wassenaar AL, Spaan WJ, Snijder EJ. An infectious arterivirus cDNA clone: identification of a replicase point mutation that abolishes discontinuous mRNA transcription. Proc Natl Acad Sci U S A. 1997;94:991–996. doi: 10.1073/pnas.94.3.991 9023370

81. Song J, Liu Y, Gao P, Hu Y, Chai Y, Zhou S, et al. Mapping the Nonstructural Protein Interaction Network of Porcine Reproductive and Respiratory Syndrome Virus. J Virol. 2018;92: e01112–01118. doi: 10.1128/JVI.01112-18 30282705

82. Du J, Ge X, Liu Y, Jiang P, Wang Z, Zhang R, et al. Targeting Swine Leukocyte Antigen Class I Molecules for Proteasomal Degradation by the nsp1alpha Replicase Protein of the Chinese Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Strain JXwn06. J Virol. 2016;90(2):682–93. doi: 10.1128/JVI.02307-15 26491168

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Hygiena a epidemiologie Infekční lékařství Laboratoř

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