Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II

Robert Sample, Locke Johnson Bryan, Scott Long, Sai Majji, Glenn Hoskins, Allan Sinning, Jake Olivier, V. Gregory Chinchar

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Frog virus 3 (FV3) is a large DNA virus that encodes ∼ 100 proteins. Although the general features of FV3 replication are known, the specific roles that most viral proteins play in the virus life cycle have not yet been elucidated. To address the question of viral gene function, antisense morpholino oligonucleotides (asMOs) were used to transiently knock-down expression of specific viral genes and thus infer their role in virus replication. We designed asMOs directed against the major capsid protein (MCP), an 18 kDa immediate-early protein (18K) that was thought to be a viral regulatory protein, and the viral homologue of the largest subunit of RNA polymerase II (vPol-IIα). All three asMOs successfully inhibited translation of the targeted protein, and two of the three asMOs resulted in marked phenotypic changes. Knock-down of the MCP resulted in a marked reduction in viral titer without a corresponding drop in the synthesis of other late viral proteins. Transmission electron microscopy (TEM) showed that in cells treated with the anti-MCP MO assembly sites were devoid of viral particles and contained numerous aberrant structures. In contrast, inhibition of 18K synthesis did not block virion formation, suggesting that the 18K protein was not essential for replication of FV3 in fathead minnow (FHM) cells. Finally, consistent with the view that late viral gene expression is catalyzed by a virus-encoded or virus-modified Pol-II-like protein, knock-down of vPol-IIα triggered a global decline in late gene expression and virus yields without affecting the synthesis of early viral genes. Collectively, these results demonstrate the utility of using asMOs to elucidate the function of FV3 proteins.

Original languageEnglish (US)
Pages (from-to)311-320
Number of pages10
JournalVirology
Volume358
Issue number2
DOIs
StatePublished - Feb 20 2007
Externally publishedYes

Fingerprint

Iridovirus
Immediate-Early Proteins
Ranavirus
Morpholinos
RNA Polymerase II
Antisense Oligonucleotides
Viral RNA
Capsid Proteins
Virus Replication
Viral Genes
Viruses
Viral Proteins
Proteins
Virion
Viral Regulatory and Accessory Proteins
Gene Expression
Cyprinidae
DNA Viruses
Protein Biosynthesis
Life Cycle Stages

Keywords

  • Antisense morpholino oligonucleotides
  • Frog virus 3
  • Ranavirus

ASJC Scopus subject areas

  • Virology

Cite this

Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II. / Sample, Robert; Bryan, Locke Johnson; Long, Scott; Majji, Sai; Hoskins, Glenn; Sinning, Allan; Olivier, Jake; Chinchar, V. Gregory.

In: Virology, Vol. 358, No. 2, 20.02.2007, p. 311-320.

Research output: Contribution to journalArticle

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