Porcine Reproductive and Respiratory Syndrome Virus Inhibits Type I Interferon Signaling by Blocking STAT1/STAT2 Nuclear Translocation

Patel, Dil V.; Nan, Yuchen; Ritthipichai, Krit; Zhu, Xiaoping; Zhang, Yan‐Jin

doi:10.1128/jvi.00655-10

Cited by 143 publications

(176 citation statements)

References 42 publications

Supporting

Mentioning

168

Contrasting

Order By: Relevance

“…It has been suggested that the PRRSV nsp1b protein may act on the IFN-b production and signalling pathways, in which it could have a direct effect on the formation of the transcription enhanceosome on the IFN-b promoter in the nucleus, as well as having an effect on the nuclear translocation of STAT1/STAT2 (Beura et al, 2010;Chen et al, 2010;Kim et al, 2010;Patel et al, 2010). The data generated from our current study consistently showed that nsp1b inhibited reporter gene mRNA expression, resulting in a strong inhibition in reporter protein synthesis.…”

Section: Discussionsupporting

confidence: 64%

“…Previous studies from our laboratory and from others have identified PRRSV nsp1b as an IFN antagonist, whereby nsp1b inhibits type I IFN activities in expression systems employing mainly reporter gene-based assays (Beura et al, 2010;Chen et al, 2010;Kim et al, 2010;Patel et al, 2010). It has been suggested that the PRRSV nsp1b protein may act on the IFN-b production and signalling pathways, in which it could have a direct effect on the formation of the transcription enhanceosome on the IFN-b promoter in the nucleus, as well as having an effect on the nuclear translocation of STAT1/STAT2 (Beura et al, 2010;Chen et al, 2010;Kim et al, 2010;Patel et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Targeted mutations in a highly conserved motif of the nsp1β protein impair the interferon antagonizing activity of porcine reproductive and respiratory syndrome virus

Lǐ

Zhu

Lawson

et al. 2013

Journal of General Virology

View full text Add to dashboard Cite

Non-structural protein 1b (nsp1b) of porcine reproductive and respiratory syndrome virus (PRRSV) contains a papain-like cysteine protease (PLPb) domain and has been identified as the main viral protein antagonizing the host innate immune response. In this study, nsp1b was determined to suppress the expression of reporter genes as well as to suppress 'self-expression' in transfected cells, and this activity appeared to be associated with its interferon (IFN) antagonist function. To knock down the effect of nsp1b on IFN activity, a panel of site-specific mutations in nsp1b was analysed. Double mutations K130A/R134A (type 1 PRRSV) or K124A/R128A (type 2 PRRSV) targeting a highly conserved motif of nsp1b, GKYLQRRLQ (in bold), impaired the ability of nsp1b to suppress IFN-b and reporter gene expression, as well as to suppress 'self-expression' in vitro. Subsequently, viable recombinant viruses vSD01-08-K130A/R134A and vSD95-21-K124A/R128A, containing double mutations in the GKYLQRRLQ motif were generated using reverse genetics. In comparison with WT viruses, these nsp1b mutants showed impaired growth ability in infected cells, but the PLPb cleavage function was not directly affected. The expression of selected innate immune genes was determined in vSD95-21-K124A/R128A mutant-infected cells. The results consistently showed that gene expression levels of IFN-a, IFN-b and IFNstimulated gene 15 were upregulated in cells that were infected with the vSD95-21-K124A/ R128A compared with that of WT virus. These data suggest that PRRSV nsp1b may selectively suppress cellular gene expression, including expression of genes involved in the host innate immune function. Modifying the key residues in the highly conserved GKYLQRRLQ motif could attenuate virus growth and improve the cellular innate immune responses.

show abstract

Section: Discussionsupporting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Targeted mutations in a highly conserved motif of the nsp1β protein impair the interferon antagonizing activity of porcine reproductive and respiratory syndrome virus

Lǐ

Zhu

Lawson

et al. 2013

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting were carried out as described previously (Patel et al, 2010;Nan et al, 2012). Briefly, after the denatured proteins had been transferred onto a polyvinylidene difluoride membrane, the membrane was blocked and probed by rabbit anti-Bax (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-Bcl-2 (Santa Cruz Biotechnology) antibodies.…”

Section: Western Blottingmentioning

confidence: 99%

“…Complementary DNA (cDNA) generation for the quantification of miRNA expression was carried out using a Hairpin-it TM miRNA RT-PCR Quantitation Kit (GenePharma, Shanghai, China) and ABM hsa-miR-152 Primers (Applied Biological Materials Inc., Richmond, BC, Canada) according to the manufacturer instructions. For cellular gene transcripts quantification, reverse transcription via AMV reverse transcriptase (Promega) was carried out using a combination of oligo dT and a random hexamer according to the manufacturer instructions, and qPCR detection with SYBR Green Mix (Life Technologies) for the targeting genes, as described previously (Patel et al, 2008;Patel et al, 2010). Transcripts of the glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) and U6 small nuclear RNA (snRNA) were also amplified from the same sample to serve as an internal control for the cellar gene or miRNA normalization, respectively.…”

Section: Reverse Transcription and Qpcrmentioning

confidence: 99%

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

Cao

et al. 2016

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Brain damage caused by perinatal asphyxia is dangerous for neonatal infants, but the mechanism by which it occurs remains elusive. In this study, microRNA-152 (miR-152) expression was induced by low oxygen levels in rat models of hypoxia brain damage, as well as in human brain microvascular endothelial cells (HBMECs) cultured in vitro. Analysis of the sequence of miR-152 revealed that the phosphatase and tensin homolog gene (PTEN) is probably the target of miR-152 both in humans and rats. When HBMECs were transfected with miR-152 mimics, PTEN expression was inhibited at both the mRNA and protein levels. Moreover, transfection with the miR-152 mimic also inhibited apoptosis induced by hypoxia. Furthermore, expression of the pro-apoptotic gene Bax was downregulated while the anti-apoptotic gene Bcl2 was upregulated after miR-152 mimic transfection. Taken together, these results indicate that miR-152 induced by hypoxia suppresses cell apoptosis and acts as a protective factor during hypoxia by repressing PTEN.

show abstract

“…SDS-PAGE and western blotting were performed as previously described (Patel et al, 2010;Nan et al, 2012). Briefly, HEK293T cells that were transfected with different plasmids were lysed in Laemmli sample buffer and subjected to SDS-PAGE.…”

Section: Western Blot Analysismentioning

confidence: 99%

Identification of a mutation in Hepatitis B virus surface antigen capable of evading ELISA screening

Yan

et al. 2016

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Hepatitis B virus (HBV) infection can cause HBVrelated cirrhosis, liver failure, and hepatocellular carcinoma. At present, a hepatitis B surface antigen (HBsAg) blood test is the primary clinical and diagnostic marker for the identification of a chronic HBV infection. In the current study, we isolated a novel HBV mutant from a chronic HBV patient, capable of causing a false negative test result for most (7 of 8) commercial HBsAg ELISA kits. DNA sequencing of the HBsAg region of this HBV mutant revealed two novel mutation sites that resulted in a Thr-to-Met substitution at amino acid (aa) position 118 and a Lys to Asn substitution at aa position 122 of HBsAg. Moreover, a mutagenesis assay showed that the aa118 (Thr to Met) mutation was the leading cause of the false negative results from the HBsAg ELISA tests. The false negative result was restored, in that the mutation was correctly detected, when the Thr at aa position 118 of this mutated HBsAg was reconstituted. In conclusion, our study revealed a novel aa118 Met mutation of HBsAg HBV that will benefit the future development of HBV diagnosis.

show abstract

Porcine Reproductive and Respiratory Syndrome Virus Inhibits Type I Interferon Signaling by Blocking STAT1/STAT2 Nuclear Translocation

Cited by 143 publications

References 42 publications

Targeted mutations in a highly conserved motif of the nsp1β protein impair the interferon antagonizing activity of porcine reproductive and respiratory syndrome virus

Targeted mutations in a highly conserved motif of the nsp1β protein impair the interferon antagonizing activity of porcine reproductive and respiratory syndrome virus

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

Identification of a mutation in Hepatitis B virus surface antigen capable of evading ELISA screening

Contact Info

Product

Resources

About