Amino acid residues in the non-structural protein 1 of porcine reproductive and respiratory syndrome virus involved in down-regulation of TNF-α expression in vitro and attenuation in vivo

Sakthivel Subramaniam, Lalit K. Beura, Byungjoon Kwon, Asit K Pattnaik, Fernando A. Osorio

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses tumor necrosis factor-alpha (TNF-α) production at both transcriptional and post-transcriptional levels by its non-structural proteins 1α and 1Β (Nsp1α and Nsp1Β). To identify the amino acid residues responsible for this activity, we generated several alanine substitution mutants of Nsp1α and Nsp1Β. Examination of the mutant proteins revealed that Nsp1α residues Gly90, Asn91, Arg97, Arg100 and Arg124 were necessary for TNF-α promoter suppression, whereas several amino acids spanning the entire Nsp1Β were found to be required for this activity. Two mutant viruses, with mutations at Nsp1α Gly90 or Nsp1Β residues 70-74, generated from infectious cDNA clones, exhibited attenuated viral replication in vitro and TNF-α was found to be up regulated in infected macrophages. In infected pigs, the Nsp1Β mutant virus was attenuated in growth. These studies provide insights into how PRRSV evades the effector mechanisms of innate immunity during infection.

Original languageEnglish (US)
Pages (from-to)241-249
Number of pages9
JournalVirology
Volume432
Issue number2
DOIs
StatePublished - Oct 25 2012

Fingerprint

Porcine respiratory and reproductive syndrome virus
Down-Regulation
Tumor Necrosis Factor-alpha
Amino Acids
Viruses
Proteins
Mutant Proteins
Innate Immunity
Alanine
Swine
Complementary DNA
Clone Cells
Macrophages
Mutation
Growth
Infection
In Vitro Techniques

Keywords

  • Immune evasion
  • Innate immunity
  • PRRSV
  • Promoter regulation
  • Reverse genetics
  • TNF-α

ASJC Scopus subject areas

  • Virology

Cite this

Amino acid residues in the non-structural protein 1 of porcine reproductive and respiratory syndrome virus involved in down-regulation of TNF-α expression in vitro and attenuation in vivo. / Subramaniam, Sakthivel; Beura, Lalit K.; Kwon, Byungjoon; Pattnaik, Asit K; Osorio, Fernando A.

In: Virology, Vol. 432, No. 2, 25.10.2012, p. 241-249.

Research output: Contribution to journalArticle

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