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Published in: Archives of Virology 7/2016

01-07-2016 | Original Article

Porcine reproductive and respiratory syndrome virus envelope (E) protein interacts with mitochondrial proteins and induces apoptosis

Authors: Sujit Pujhari, Alexander N. Zakhartchouk

Published in: Archives of Virology | Issue 7/2016

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Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant economic losses for the swine industry worldwide. The PRRSV E protein, encoded by ORF 2b, is one of the non-glycosylated minor structural proteins. In this study, we present evidence for the interaction of the E protein with mitochondrial proteins ATP5A (part of ATP synthase complex), prohibitin, and ADP/ATP translocase. We additionally demonstrate partial mitochondrial localization of the E protein in transfected cells. To functionally investigate these interactions, we infected MARC-145 cells with PRRSV or alphavirus replicon particles (VRPs) expressing PRRSV E protein. In infected cells, production of ATP was significantly reduced. The E protein also induced apoptosis by activating caspase-3, which results in PARP cleavage. Taken together, these data suggest that the PRRSV E protein interacts with mitochondrial proteins and induces apoptosis by inhibiting ATP production.
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Literature
1.
go back to reference Abu-Hamad S, Sivan S, Shoshan-Barmatz V (2006) The expression level of the voltage-dependent anion channel controls life and death of the cell. Proceedings of the National Academy of Sciences of the United States of America 103:5787–5792CrossRefPubMedPubMedCentral Abu-Hamad S, Sivan S, Shoshan-Barmatz V (2006) The expression level of the voltage-dependent anion channel controls life and death of the cell. Proceedings of the National Academy of Sciences of the United States of America 103:5787–5792CrossRefPubMedPubMedCentral
3.
go back to reference Atkins GJ, Fleeton MN, Sheahan BJ (2008) Therapeutic and prophylactic applications of alphavirus vectors. Expert Rev Mole Med 10:e33CrossRef Atkins GJ, Fleeton MN, Sheahan BJ (2008) Therapeutic and prophylactic applications of alphavirus vectors. Expert Rev Mole Med 10:e33CrossRef
4.
go back to reference Aweya JJ, Mak TM, Lim SG, Tan YJ (2013) The p7 protein of the hepatitis C virus induces cell death differently from the influenza A virus viroporin M2. Virus Res 172:24–34CrossRefPubMed Aweya JJ, Mak TM, Lim SG, Tan YJ (2013) The p7 protein of the hepatitis C virus induces cell death differently from the influenza A virus viroporin M2. Virus Res 172:24–34CrossRefPubMed
5.
go back to reference Beauchamp E, Tekpli X, Marteil G, Lagadic-Gossmann D, Legrand P, Rioux V (2009) N-Myristoylation targets dihydroceramide Delta4-desaturase 1 to mitochondria: partial involvement in the apoptotic effect of myristic acid. Biochimie 91:1411–1419CrossRefPubMed Beauchamp E, Tekpli X, Marteil G, Lagadic-Gossmann D, Legrand P, Rioux V (2009) N-Myristoylation targets dihydroceramide Delta4-desaturase 1 to mitochondria: partial involvement in the apoptotic effect of myristic acid. Biochimie 91:1411–1419CrossRefPubMed
6.
go back to reference Bradbury DA, Simmons TD, Slater KJ, Crouch SP (2000) Measurement of the ADP:ATP ratio in human leukaemic cell lines can be used as an indicator of cell viability, necrosis and apoptosis. J Immunol Methods 240:79–92CrossRefPubMed Bradbury DA, Simmons TD, Slater KJ, Crouch SP (2000) Measurement of the ADP:ATP ratio in human leukaemic cell lines can be used as an indicator of cell viability, necrosis and apoptosis. J Immunol Methods 240:79–92CrossRefPubMed
7.
go back to reference Butler JE, Lager KM, Golde W, Faaberg KS, Sinkora M, Loving C, Zhang YI (2014) Porcine reproductive and respiratory syndrome (PRRS): an immune dysregulatory pandemic. Immunol Res 59:81–108CrossRefPubMed Butler JE, Lager KM, Golde W, Faaberg KS, Sinkora M, Loving C, Zhang YI (2014) Porcine reproductive and respiratory syndrome (PRRS): an immune dysregulatory pandemic. Immunol Res 59:81–108CrossRefPubMed
8.
go back to reference Colombo S, Longhi R, Alcaro S, Ortuso F, Sprocati T, Flora A, Borgese N (2005) N-myristoylation determines dual targeting of mammalian NADH-cytochrome b5 reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning. J Cell Biol 168:735–745CrossRefPubMedPubMedCentral Colombo S, Longhi R, Alcaro S, Ortuso F, Sprocati T, Flora A, Borgese N (2005) N-myristoylation determines dual targeting of mammalian NADH-cytochrome b5 reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning. J Cell Biol 168:735–745CrossRefPubMedPubMedCentral
9.
go back to reference Du Y, Zuckermann FA, Yoo D (2010) Myristoylation of the small envelope protein of porcine reproductive and respiratory syndrome virus is non-essential for virus infectivity but promotes its growth. Virus research 147:294–299CrossRefPubMed Du Y, Zuckermann FA, Yoo D (2010) Myristoylation of the small envelope protein of porcine reproductive and respiratory syndrome virus is non-essential for virus infectivity but promotes its growth. Virus research 147:294–299CrossRefPubMed
10.
go back to reference Eguchi Y, Shimizu S, Tsujimoto Y (1997) Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res 57:1835–1840PubMed Eguchi Y, Shimizu S, Tsujimoto Y (1997) Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res 57:1835–1840PubMed
11.
go back to reference Fernandez A, Suarez P, Castro JM, Tabares E, Diaz-Guerra M (2002) Characterization of regions in the GP5 protein of porcine reproductive and respiratory syndrome virus required to induce apoptotic cell death. Virus Res 83:103–118CrossRefPubMed Fernandez A, Suarez P, Castro JM, Tabares E, Diaz-Guerra M (2002) Characterization of regions in the GP5 protein of porcine reproductive and respiratory syndrome virus required to induce apoptotic cell death. Virus Res 83:103–118CrossRefPubMed
12.
go back to reference Griffin SD, Harvey R, Clarke DS, Barclay WS, Harris M, Rowlands DJ (2004) A conserved basic loop in hepatitis C virus p7 protein is required for amantadine-sensitive ion channel activity in mammalian cells but is dispensable for localization to mitochondria. J General Virol 85:451–461CrossRef Griffin SD, Harvey R, Clarke DS, Barclay WS, Harris M, Rowlands DJ (2004) A conserved basic loop in hepatitis C virus p7 protein is required for amantadine-sensitive ion channel activity in mammalian cells but is dispensable for localization to mitochondria. J General Virol 85:451–461CrossRef
13.
go back to reference Jacotot E, Ferri KF, El Hamel C, Brenner C, Druillennec S, Hoebeke J, Rustin P, Metivier D, Lenoir C, Geuskens M, Vieira HL, Loeffler M, Belzacq AS, Briand JP, Zamzami N, Edelman L, Xie ZH, Reed JC, Roques BP, Kroemer G (2001) Control of mitochondrial membrane permeabilization by adenine nucleotide translocator interacting with HIV-1 viral protein rR and Bcl-2. J Exp Med 193:509–519CrossRefPubMedPubMedCentral Jacotot E, Ferri KF, El Hamel C, Brenner C, Druillennec S, Hoebeke J, Rustin P, Metivier D, Lenoir C, Geuskens M, Vieira HL, Loeffler M, Belzacq AS, Briand JP, Zamzami N, Edelman L, Xie ZH, Reed JC, Roques BP, Kroemer G (2001) Control of mitochondrial membrane permeabilization by adenine nucleotide translocator interacting with HIV-1 viral protein rR and Bcl-2. J Exp Med 193:509–519CrossRefPubMedPubMedCentral
14.
go back to reference Kim HS, Kwang J, Yoon IJ, Joo HS, Frey ML (1993) Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line. Arch Virol 133:477–483CrossRefPubMed Kim HS, Kwang J, Yoon IJ, Joo HS, Frey ML (1993) Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line. Arch Virol 133:477–483CrossRefPubMed
15.
go back to reference Lee C, Rogan D, Erickson L, Zhang J, Yoo D (2004) Characterization of the porcine reproductive and respiratory syndrome virus glycoprotein 5 (GP5) in stably expressing cells. Virus Res 104:33–38CrossRefPubMed Lee C, Rogan D, Erickson L, Zhang J, Yoo D (2004) Characterization of the porcine reproductive and respiratory syndrome virus glycoprotein 5 (GP5) in stably expressing cells. Virus Res 104:33–38CrossRefPubMed
16.
go back to reference Lee C, Yoo D (2006) The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties. Virol 355:30–43CrossRef Lee C, Yoo D (2006) The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties. Virol 355:30–43CrossRef
17.
go back to reference Lee SM, Kleiboeker SB (2007) Porcine reproductive and respiratory syndrome virus induces apoptosis through a mitochondria-mediated pathway. Virology 365:419–434CrossRefPubMed Lee SM, Kleiboeker SB (2007) Porcine reproductive and respiratory syndrome virus induces apoptosis through a mitochondria-mediated pathway. Virology 365:419–434CrossRefPubMed
18.
go back to reference Ma Z, Wang Y, Zhao H, Xu AT, Tang J, Feng WH (2013) Porcine reproductive and respiratory syndrome virus nonstructural protein 4 induces apoptosis dependent on its 3C-like serine protease activity. PloS One 8:e69387CrossRefPubMedPubMedCentral Ma Z, Wang Y, Zhao H, Xu AT, Tang J, Feng WH (2013) Porcine reproductive and respiratory syndrome virus nonstructural protein 4 induces apoptosis dependent on its 3C-like serine protease activity. PloS One 8:e69387CrossRefPubMedPubMedCentral
19.
go back to reference Mu Y, Li L, Zhang B, Huang B, Gao J, Wang X, Wang C, Xiao S, Zhao Q, Sun Y, Zhang G, Hiscox JA, Zhou EM (2015) Glycoprotein 5 of porcine reproductive and respiratory syndrome virus strain SD16 inhibits viral replication and causes G2/M cell cycle arrest, but does not induce cellular apoptosis in Marc-145 cells. Virology 484:136–145CrossRefPubMed Mu Y, Li L, Zhang B, Huang B, Gao J, Wang X, Wang C, Xiao S, Zhao Q, Sun Y, Zhang G, Hiscox JA, Zhou EM (2015) Glycoprotein 5 of porcine reproductive and respiratory syndrome virus strain SD16 inhibits viral replication and causes G2/M cell cycle arrest, but does not induce cellular apoptosis in Marc-145 cells. Virology 484:136–145CrossRefPubMed
20.
go back to reference Nelsen CJ, Murtaugh MP, Faaberg KS (1999) Porcine reproductive and respiratory syndrome virus comparison: divergent evolution on two continents. J Virol 73:270–280PubMedPubMedCentral Nelsen CJ, Murtaugh MP, Faaberg KS (1999) Porcine reproductive and respiratory syndrome virus comparison: divergent evolution on two continents. J Virol 73:270–280PubMedPubMedCentral
21.
go back to reference Nijtmans LG, de Jong L, Artal Sanz M, Coates PJ, Berden JA, Back JW, Muijsers AO, van der Spek H, Grivell LA (2000) Prohibitins act as a membrane-bound chaperone for the stabilization of mitochondrial proteins. EMBO J 19:2444–2451CrossRefPubMedPubMedCentral Nijtmans LG, de Jong L, Artal Sanz M, Coates PJ, Berden JA, Back JW, Muijsers AO, van der Spek H, Grivell LA (2000) Prohibitins act as a membrane-bound chaperone for the stabilization of mitochondrial proteins. EMBO J 19:2444–2451CrossRefPubMedPubMedCentral
22.
go back to reference Rahmani Z, Huh KW, Lasher R, Siddiqui A (2000) Hepatitis B virus X protein colocalizes to mitochondria with a human voltage-dependent anion channel, HVDAC3, and alters its transmembrane potential. J Virol 74:2840–2846CrossRefPubMedPubMedCentral Rahmani Z, Huh KW, Lasher R, Siddiqui A (2000) Hepatitis B virus X protein colocalizes to mitochondria with a human voltage-dependent anion channel, HVDAC3, and alters its transmembrane potential. J Virol 74:2840–2846CrossRefPubMedPubMedCentral
23.
go back to reference Shoshan-Barmatz V, Israelson A, Brdiczka D, Sheu SS (2006) The voltage-dependent anion channel (VDAC): function in intracellular signalling, cell life and cell death. Curr Pharm Des 12:2249–2270CrossRefPubMed Shoshan-Barmatz V, Israelson A, Brdiczka D, Sheu SS (2006) The voltage-dependent anion channel (VDAC): function in intracellular signalling, cell life and cell death. Curr Pharm Des 12:2249–2270CrossRefPubMed
24.
go back to reference Sirinarumitr T, Zhang Y, Kluge JP, Halbur PG, Paul PS (1998) A pneumo-virulent United States isolate of porcine reproductive and respiratory syndrome virus induces apoptosis in bystander cells both in vitro and in vivo. J General Virol 79(Pt 12):2989–2995CrossRef Sirinarumitr T, Zhang Y, Kluge JP, Halbur PG, Paul PS (1998) A pneumo-virulent United States isolate of porcine reproductive and respiratory syndrome virus induces apoptosis in bystander cells both in vitro and in vivo. J General Virol 79(Pt 12):2989–2995CrossRef
25.
go back to reference Snijder EJ, Kikkert M, Fang Y (2013) Arterivirus molecular biology and pathogenesis. J General Virol 94:2141–2163CrossRef Snijder EJ, Kikkert M, Fang Y (2013) Arterivirus molecular biology and pathogenesis. J General Virol 94:2141–2163CrossRef
26.
go back to reference Suarez P, Diaz-Guerra M, Prieto C, Esteban M, Castro JM, Nieto A, Ortin J (1996) Open reading frame 5 of porcine reproductive and respiratory syndrome virus as a cause of virus-induced apoptosis. J Virol 70:2876–2882PubMedPubMedCentral Suarez P, Diaz-Guerra M, Prieto C, Esteban M, Castro JM, Nieto A, Ortin J (1996) Open reading frame 5 of porcine reproductive and respiratory syndrome virus as a cause of virus-induced apoptosis. J Virol 70:2876–2882PubMedPubMedCentral
27.
go back to reference Veit M, Matczuk AK, Sinhadri BC, Krause E, Thaa B (2014) Membrane proteins of arterivirus particles: structure, topology, processing and function. Virus Res 194:16–36CrossRefPubMed Veit M, Matczuk AK, Sinhadri BC, Krause E, Thaa B (2014) Membrane proteins of arterivirus particles: structure, topology, processing and function. Virus Res 194:16–36CrossRefPubMed
28.
go back to reference Volkova E, Gorchakov R, Frolov I (2006) The efficient packaging of Venezuelan equine encephalitis virus-specific RNAs into viral particles is determined by nsP1-3 synthesis. Virology 344:315–327CrossRefPubMed Volkova E, Gorchakov R, Frolov I (2006) The efficient packaging of Venezuelan equine encephalitis virus-specific RNAs into viral particles is determined by nsP1-3 synthesis. Virology 344:315–327CrossRefPubMed
29.
go back to reference Wang X, Li C, Zhou L, Zhang N, Ge X, Guo X, Yang H (2014) Porcine reproductive and respiratory syndrome virus counteracts the porcine intrinsic virus restriction factors-IFITM1 and Tetherin in MARC-145 cells. Virus Res 191:92–100CrossRefPubMed Wang X, Li C, Zhou L, Zhang N, Ge X, Guo X, Yang H (2014) Porcine reproductive and respiratory syndrome virus counteracts the porcine intrinsic virus restriction factors-IFITM1 and Tetherin in MARC-145 cells. Virus Res 191:92–100CrossRefPubMed
30.
go back to reference Wissink EH, Kroese MV, van Wijk HA, Rijsewijk FA, Meulenberg JJ, Rottier PJ (2005) Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus. J Virol 79:12495–12506CrossRefPubMedPubMedCentral Wissink EH, Kroese MV, van Wijk HA, Rijsewijk FA, Meulenberg JJ, Rottier PJ (2005) Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus. J Virol 79:12495–12506CrossRefPubMedPubMedCentral
31.
go back to reference Wu WH, Fang Y, Farwell R, Steffen-Bien M, Rowland RR, Christopher-Hennings J, Nelson EA (2001) A 10-kDa structural protein of porcine reproductive and respiratory syndrome virus encoded by ORF2b. Virology 287:183–191CrossRefPubMed Wu WH, Fang Y, Farwell R, Steffen-Bien M, Rowland RR, Christopher-Hennings J, Nelson EA (2001) A 10-kDa structural protein of porcine reproductive and respiratory syndrome virus encoded by ORF2b. Virology 287:183–191CrossRefPubMed
32.
go back to reference Wu WH, Fang Y, Rowland RR, Lawson SR, Christopher-Hennings J, Yoon KJ, Nelson EA (2005) The 2b protein as a minor structural component of PRRSV. Virus Res 114:177–181CrossRefPubMed Wu WH, Fang Y, Rowland RR, Lawson SR, Christopher-Hennings J, Yoon KJ, Nelson EA (2005) The 2b protein as a minor structural component of PRRSV. Virus Res 114:177–181CrossRefPubMed
33.
go back to reference Yu M, Liu X, Sun L, Chen C, Ma G, Kitamura Y, Gao GF, Liu W (2010) Subcellular localization and topology of porcine reproductive and respiratory syndrome virus E protein. Virus Res 152:104–114CrossRefPubMed Yu M, Liu X, Sun L, Chen C, Ma G, Kitamura Y, Gao GF, Liu W (2010) Subcellular localization and topology of porcine reproductive and respiratory syndrome virus E protein. Virus Res 152:104–114CrossRefPubMed
34.
go back to reference Zamarin D, Garcia-Sastre A, Xiao X, Wang R, Palese P (2005) Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS Pathog 1:e4CrossRefPubMedPubMedCentral Zamarin D, Garcia-Sastre A, Xiao X, Wang R, Palese P (2005) Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS Pathog 1:e4CrossRefPubMedPubMedCentral
35.
go back to reference Zhang K, Hou Q, Zhong Z, Li X, Chen H, Li W, Wen J, Wang L, Liu W, Zhong F (2013) Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E. Virology 442:156–162CrossRefPubMed Zhang K, Hou Q, Zhong Z, Li X, Chen H, Li W, Wen J, Wang L, Liu W, Zhong F (2013) Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E. Virology 442:156–162CrossRefPubMed
Metadata
Title
Porcine reproductive and respiratory syndrome virus envelope (E) protein interacts with mitochondrial proteins and induces apoptosis
Authors
Sujit Pujhari
Alexander N. Zakhartchouk
Publication date
01-07-2016
Publisher
Springer Vienna
Published in
Archives of Virology / Issue 7/2016
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-016-2845-4

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