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Published in: Archives of Virology 2/2016

01-02-2016 | Original Article

Protein kinase R (PKR) plays a pro-viral role in porcine reproductive and respiratory syndrome virus (PRRSV) replication by modulating viral gene transcription

Authors: Xiuqing Wang, Hanmo Zhang, Alex M. Abel, Eric Nelson

Published in: Archives of Virology | Issue 2/2016

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Abstract

Protein kinase R (PKR) is involved in apoptotic cell death and antiviral activities in response to many virus infections. To reveal the role of PKR in the replication of porcine reproductive and respiratory syndrome virus (PRRSV), we first examined the kinetics of PKR phosphorylation during PRRSV infection. The results showed that PRRSV transiently activates PKR at 12 and 24 h postinfection. Surprisingly, eIF-2α, the well-known downstream target of PKR, was significantly phosphorylated compared to mock-infected cells only at 48 and 72 h postinfection. Reduced viral gene transcription, viral protein synthesis, and virus titer were detected in cells transfected with PKR silencing RNA prior to PRRSV infection compared to control silencing RNA transfected cells, indicating a role of PKR in facilitating virus replication. Overall, our data suggest that PKR is not a major contributor to the phosphorylation of eIF-2α during PRRSV infection, but it plays a pro-viral role in PRRSV replication by modulating primarily viral gene transcription.
Literature
1.
go back to reference Adair R, Patel AH, Corless L, Griffin S, Rowlands DJ, McCormick CJ (2009) Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA. J Gen Virol 90:833–842CrossRefPubMed Adair R, Patel AH, Corless L, Griffin S, Rowlands DJ, McCormick CJ (2009) Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA. J Gen Virol 90:833–842CrossRefPubMed
2.
go back to reference Arnaud N, Dabo S, Maillard P, Budkowska A, Kalliampakou KI, Mavromara P, Garcin D, Hugon J, Gatignol A, Akazawa D, Wakita T, Meurs EF (2010) Hepatitis C virus controls interferon production through PKR activation. PLoS One 5:e10575PubMedCentralCrossRefPubMed Arnaud N, Dabo S, Maillard P, Budkowska A, Kalliampakou KI, Mavromara P, Garcin D, Hugon J, Gatignol A, Akazawa D, Wakita T, Meurs EF (2010) Hepatitis C virus controls interferon production through PKR activation. PLoS One 5:e10575PubMedCentralCrossRefPubMed
3.
go back to reference Barry G, Breakwell L, Fragkoudis R, Attarzadeh-Yazdi G, Rodriguez-Andres J, Kohl A, Fazakerley JK (2009) PKR acts early in infection to suppress Semliki Forest virus production and strongly enhances the type I interferon response. J Gen Virol 90:1382–1391PubMedCentralCrossRefPubMed Barry G, Breakwell L, Fragkoudis R, Attarzadeh-Yazdi G, Rodriguez-Andres J, Kohl A, Fazakerley JK (2009) PKR acts early in infection to suppress Semliki Forest virus production and strongly enhances the type I interferon response. J Gen Virol 90:1382–1391PubMedCentralCrossRefPubMed
4.
go back to reference Cai R, Carpick B, Chun RF, Jeang KT, Williams BR (2000) HIV-I TAT inhibits PKR activity by both RNA-dependent and RNA-independent mechanisms. Arch Biochem Biophys 373:361–367CrossRefPubMed Cai R, Carpick B, Chun RF, Jeang KT, Williams BR (2000) HIV-I TAT inhibits PKR activity by both RNA-dependent and RNA-independent mechanisms. Arch Biochem Biophys 373:361–367CrossRefPubMed
5.
go back to reference Dauber B, Martinez-Sobrido L, Schneider J, Hai R, Waibler Z, Kalinke U, Garcia-Sastre A, Wolff T (2009) Influenza B virus ribonucleoprotein is a potent activator of the antiviral kinase PKR. PLoS Pathog 5:e1000473PubMedCentralCrossRefPubMed Dauber B, Martinez-Sobrido L, Schneider J, Hai R, Waibler Z, Kalinke U, Garcia-Sastre A, Wolff T (2009) Influenza B virus ribonucleoprotein is a potent activator of the antiviral kinase PKR. PLoS Pathog 5:e1000473PubMedCentralCrossRefPubMed
6.
go back to reference Diamond MS, Harris E (2001) Interferon inhibits dengue virus infection by preventing translation of viral RNA through a PKR-independent mechanism. Virology 289:297–311CrossRefPubMed Diamond MS, Harris E (2001) Interferon inhibits dengue virus infection by preventing translation of viral RNA through a PKR-independent mechanism. Virology 289:297–311CrossRefPubMed
7.
go back to reference Garaigorta U, Chisari FV (2009) Hepatitis C virus blocks interferon effector function by inducing protein kinase R phosphorylation. Cell Host Microbe 6:513–522PubMedCentralCrossRefPubMed Garaigorta U, Chisari FV (2009) Hepatitis C virus blocks interferon effector function by inducing protein kinase R phosphorylation. Cell Host Microbe 6:513–522PubMedCentralCrossRefPubMed
8.
go back to reference Garaigorta U, Heim MH, Boyd B, Wieland S, Chisari FV (2012) Hepatitis C virus (HCV) induces formation of stress granules whose proteins regulate HCV RNA replication and virus assembly and egress. J Virol 86:11043–11056PubMedCentralCrossRefPubMed Garaigorta U, Heim MH, Boyd B, Wieland S, Chisari FV (2012) Hepatitis C virus (HCV) induces formation of stress granules whose proteins regulate HCV RNA replication and virus assembly and egress. J Virol 86:11043–11056PubMedCentralCrossRefPubMed
9.
go back to reference Hakki M, Marshall EE, De Niro KL, Geballe AP (2006) Binding and nuclear relocalization of protein kinase R by human cytomegalovirus TRS1. J Virol 80:11817–11826PubMedCentralCrossRefPubMed Hakki M, Marshall EE, De Niro KL, Geballe AP (2006) Binding and nuclear relocalization of protein kinase R by human cytomegalovirus TRS1. J Virol 80:11817–11826PubMedCentralCrossRefPubMed
11.
go back to reference Huo Y, Fan L, Yin S, Dong Y, Guo X, Yang H, Hu H (2013) Involvement of unfolded protein response, p53 and Akt in modulation of porcine reproductive and respiratory syndrome virus-mediated JNK activation. Virology 444:233–240CrossRefPubMed Huo Y, Fan L, Yin S, Dong Y, Guo X, Yang H, Hu H (2013) Involvement of unfolded protein response, p53 and Akt in modulation of porcine reproductive and respiratory syndrome virus-mediated JNK activation. Virology 444:233–240CrossRefPubMed
12.
go back to reference Irving AT, Wang D, Vasilevski O, Latchoumanin O, Kozer N, Clayton AH, Szczepny A, Morimoto H, Xu D, Williams BR, Sadler AJ (2012) Regulation of actin dynamics by protein kinase R control of gelsolin enforces basal innate immune defense. Immunity 36:795–806CrossRefPubMed Irving AT, Wang D, Vasilevski O, Latchoumanin O, Kozer N, Clayton AH, Szczepny A, Morimoto H, Xu D, Williams BR, Sadler AJ (2012) Regulation of actin dynamics by protein kinase R control of gelsolin enforces basal innate immune defense. Immunity 36:795–806CrossRefPubMed
13.
go back to reference Krishnamoorthy J, Mounir Z, Raven JF, Koromilas AE (2008) The eIF2alpha kinases inhibit vesicular stomatitis virus replication independently of eIF2alpha phosphorylation. Cell Cycle 7:2346–2351CrossRefPubMed Krishnamoorthy J, Mounir Z, Raven JF, Koromilas AE (2008) The eIF2alpha kinases inhibit vesicular stomatitis virus replication independently of eIF2alpha phosphorylation. Cell Cycle 7:2346–2351CrossRefPubMed
14.
go back to reference Leib DA, Machalek MA, Williams BR, Silverman RH, Virgin HW (2000) Specific phenotypic restoration of an attenuated virus by knockout of a host resistance gene. Proc Natl Acad Sci USA 97:6097–6101PubMedCentralCrossRefPubMed Leib DA, Machalek MA, Williams BR, Silverman RH, Virgin HW (2000) Specific phenotypic restoration of an attenuated virus by knockout of a host resistance gene. Proc Natl Acad Sci USA 97:6097–6101PubMedCentralCrossRefPubMed
15.
go back to reference Li S, Min JY, Krug RM, Sen GC (2006) Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA. Virology 349:13–21CrossRefPubMed Li S, Min JY, Krug RM, Sen GC (2006) Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA. Virology 349:13–21CrossRefPubMed
16.
go back to reference Li Z, Okonski KM, Samuel CE (2012) Adenosine deaminase acting on RNA 1 (ADAR1) suppresses the induction of interferon by measles virus. J Virol 86:3787–3794PubMedCentralCrossRefPubMed Li Z, Okonski KM, Samuel CE (2012) Adenosine deaminase acting on RNA 1 (ADAR1) suppresses the induction of interferon by measles virus. J Virol 86:3787–3794PubMedCentralCrossRefPubMed
17.
go back to reference Lindquist ME, Mainou BA, Dermody TS, Crowe JE Jr (2011) Activation of protein kinase R is required for induction of stress granules by respiratory syncytial virus but dispensable for viral replication. Virology 413:103–110PubMedCentralCrossRefPubMed Lindquist ME, Mainou BA, Dermody TS, Crowe JE Jr (2011) Activation of protein kinase R is required for induction of stress granules by respiratory syncytial virus but dispensable for viral replication. Virology 413:103–110PubMedCentralCrossRefPubMed
19.
go back to reference Loving CL, Brockmeier SL, Ma W, Richt JA, Sacco RE (2006) Innate cytokine responses in porcine macrophage populations: evidence for differential recognition of double-stranded RNA. J Immunol 177:8432–8439CrossRefPubMed Loving CL, Brockmeier SL, Ma W, Richt JA, Sacco RE (2006) Innate cytokine responses in porcine macrophage populations: evidence for differential recognition of double-stranded RNA. J Immunol 177:8432–8439CrossRefPubMed
20.
go back to reference Marciniak SJ, Yun CY, Oyadomari S, Novoa I, Zhang Y, Jungreis R, Nagata K, Harding HP, Ron D (2004) CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev 18:3066–3077PubMedCentralCrossRefPubMed Marciniak SJ, Yun CY, Oyadomari S, Novoa I, Zhang Y, Jungreis R, Nagata K, Harding HP, Ron D (2004) CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev 18:3066–3077PubMedCentralCrossRefPubMed
21.
go back to reference McAllister CS, Taghavi N, Samuel CE (2012) Protein kinase PKR amplification of interferon beta induction occurs through initiation factor eIF-2alpha-mediated translational control. J Biol Chem 287:36384–36392PubMedCentralCrossRefPubMed McAllister CS, Taghavi N, Samuel CE (2012) Protein kinase PKR amplification of interferon beta induction occurs through initiation factor eIF-2alpha-mediated translational control. J Biol Chem 287:36384–36392PubMedCentralCrossRefPubMed
22.
go back to reference Nishitoh H (2012) CHOP is a multifunctional transcription factor in the ER stress response. J Biochem 151:217–219CrossRefPubMed Nishitoh H (2012) CHOP is a multifunctional transcription factor in the ER stress response. J Biochem 151:217–219CrossRefPubMed
23.
go back to reference Overend C, Mitchell R, He D, Rompato G, Grubman MJ, Garmendia AE (2007) Recombinant swine beta interferon protects swine alveolar macrophages and MARC-145 cells from infection with Porcine reproductive and respiratory syndrome virus. J Gen Virol 88:925–931CrossRefPubMed Overend C, Mitchell R, He D, Rompato G, Grubman MJ, Garmendia AE (2007) Recombinant swine beta interferon protects swine alveolar macrophages and MARC-145 cells from infection with Porcine reproductive and respiratory syndrome virus. J Gen Virol 88:925–931CrossRefPubMed
24.
go back to reference Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389CrossRefPubMed Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389CrossRefPubMed
25.
go back to reference Posti K, Leinonen S, Tetri S, Kottari S, Viitala P, Pelkonen O, Raunio H (1999) Modulation of murine phenobarbital-inducible CYP2A5, CYP2B10 and CYP1A enzymes by inhibitors of protein kinases and phosphatases. Eur J Biochem FEBS 264:19–26CrossRef Posti K, Leinonen S, Tetri S, Kottari S, Viitala P, Pelkonen O, Raunio H (1999) Modulation of murine phenobarbital-inducible CYP2A5, CYP2B10 and CYP1A enzymes by inhibitors of protein kinases and phosphatases. Eur J Biochem FEBS 264:19–26CrossRef
26.
go back to reference Rivas-Estilla AM, Svitkin Y, Lopez Lastra M, Hatzoglou M, Sherker A, Koromilas AE (2002) PKR-dependent mechanisms of gene expression from a subgenomic hepatitis C virus clone. J Virol 76:10637–10653PubMedCentralCrossRefPubMed Rivas-Estilla AM, Svitkin Y, Lopez Lastra M, Hatzoglou M, Sherker A, Koromilas AE (2002) PKR-dependent mechanisms of gene expression from a subgenomic hepatitis C virus clone. J Virol 76:10637–10653PubMedCentralCrossRefPubMed
27.
go back to reference Rojas M, Arias CF, Lopez S (2010) Protein kinase R is responsible for the phosphorylation of eIF2alpha in rotavirus infection. J Virol 84:10457–10466PubMedCentralCrossRefPubMed Rojas M, Arias CF, Lopez S (2010) Protein kinase R is responsible for the phosphorylation of eIF2alpha in rotavirus infection. J Virol 84:10457–10466PubMedCentralCrossRefPubMed
28.
go back to reference Rowland RR, Robinson B, Stefanick J, Kim TS, Guanghua L, Lawson SR, Benfield DA (2001) Inhibition of porcine reproductive and respiratory syndrome virus by interferon-gamma and recovery of virus replication with 2-aminopurine. Arch Virol 146:539–555CrossRefPubMed Rowland RR, Robinson B, Stefanick J, Kim TS, Guanghua L, Lawson SR, Benfield DA (2001) Inhibition of porcine reproductive and respiratory syndrome virus by interferon-gamma and recovery of virus replication with 2-aminopurine. Arch Virol 146:539–555CrossRefPubMed
29.
go back to reference Stojdl DF, Abraham N, Knowles S, Marius R, Brasey A, Lichty BD, Brown EG, Sonenberg N, Bell JC (2000) The murine double-stranded RNA-dependent protein kinase PKR is required for resistance to vesicular stomatitis virus. J Virol 74:9580–9585PubMedCentralCrossRefPubMed Stojdl DF, Abraham N, Knowles S, Marius R, Brasey A, Lichty BD, Brown EG, Sonenberg N, Bell JC (2000) The murine double-stranded RNA-dependent protein kinase PKR is required for resistance to vesicular stomatitis virus. J Virol 74:9580–9585PubMedCentralCrossRefPubMed
30.
go back to reference Wang X, Eaton M, Mayer M, Li H, He D, Nelson E, Christopher-Hennings J (2007) Porcine reproductive and respiratory syndrome virus productively infects monocyte-derived dendritic cells and compromises their antigen-presenting ability. Arch Virol 152:289–303CrossRefPubMed Wang X, Eaton M, Mayer M, Li H, He D, Nelson E, Christopher-Hennings J (2007) Porcine reproductive and respiratory syndrome virus productively infects monocyte-derived dendritic cells and compromises their antigen-presenting ability. Arch Virol 152:289–303CrossRefPubMed
31.
go back to reference Wang X, Liao Y, Yap PL, Png KJ, Tam JP, Liu DX (2009) Inhibition of protein kinase R activation and upregulation of GADD34 expression play a synergistic role in facilitating coronavirus replication by maintaining de novo protein synthesis in virus-infected cells. J Virol 83:12462–12472PubMedCentralCrossRefPubMed Wang X, Liao Y, Yap PL, Png KJ, Tam JP, Liu DX (2009) Inhibition of protein kinase R activation and upregulation of GADD34 expression play a synergistic role in facilitating coronavirus replication by maintaining de novo protein synthesis in virus-infected cells. J Virol 83:12462–12472PubMedCentralCrossRefPubMed
32.
go back to reference Wang X, Zhang H, Abel AM, Young AJ, Xie L, Xie Z (2014) Role of phosphatidylinositol 3-kinase (PI3K) and Akt1 kinase in porcine reproductive and respiratory syndrome virus (PRRSV) replication. Arch Virol 159:2091–2096CrossRefPubMed Wang X, Zhang H, Abel AM, Young AJ, Xie L, Xie Z (2014) Role of phosphatidylinositol 3-kinase (PI3K) and Akt1 kinase in porcine reproductive and respiratory syndrome virus (PRRSV) replication. Arch Virol 159:2091–2096CrossRefPubMed
34.
go back to reference Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K (2001) XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell 107:881–891CrossRefPubMed Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K (2001) XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell 107:881–891CrossRefPubMed
35.
go back to reference Zhang H, Guo X, Nelson E, Christopher-Hennings J, Wang X (2012) Porcine reproductive and respiratory syndrome virus activates the transcription of interferon alpha/beta (IFN-alpha/beta) in monocyte-derived dendritic cells (Mo-DC). Vet Microbiol 159:494–498CrossRefPubMed Zhang H, Guo X, Nelson E, Christopher-Hennings J, Wang X (2012) Porcine reproductive and respiratory syndrome virus activates the transcription of interferon alpha/beta (IFN-alpha/beta) in monocyte-derived dendritic cells (Mo-DC). Vet Microbiol 159:494–498CrossRefPubMed
36.
go back to reference Zhang P, Samuel CE (2007) Protein kinase PKR plays a stimulus- and virus-dependent role in apoptotic death and virus multiplication in human cells. J Virol 81:8192–8200PubMedCentralCrossRefPubMed Zhang P, Samuel CE (2007) Protein kinase PKR plays a stimulus- and virus-dependent role in apoptotic death and virus multiplication in human cells. J Virol 81:8192–8200PubMedCentralCrossRefPubMed
37.
go back to reference Zhang P, Jacobs BL, Samuel CE (2008) Loss of protein kinase PKR expression in human HeLa cells complements the vaccinia virus E3L deletion mutant phenotype by restoration of viral protein synthesis. J Virol 82:840–848PubMedCentralCrossRefPubMed Zhang P, Jacobs BL, Samuel CE (2008) Loss of protein kinase PKR expression in human HeLa cells complements the vaccinia virus E3L deletion mutant phenotype by restoration of viral protein synthesis. J Virol 82:840–848PubMedCentralCrossRefPubMed
Metadata
Title
Protein kinase R (PKR) plays a pro-viral role in porcine reproductive and respiratory syndrome virus (PRRSV) replication by modulating viral gene transcription
Authors
Xiuqing Wang
Hanmo Zhang
Alex M. Abel
Eric Nelson
Publication date
01-02-2016
Publisher
Springer Vienna
Published in
Archives of Virology / Issue 2/2016
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-015-2671-0

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