Expression of the insect parvovirus GmDNV in vivo: The structural and nonstructural proteins are encoded by opposite DNA strands

Orit Gross, Jacov Tal

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

The nucleotide sequence of GmDNV, an insect parvovirus, reveals large open reading frames (ORFs) on both strands of the viral replicative form DNA. Previously, we identified two viral transcripts within the polyadenylated RNA fraction of infected host larvae (Gross et al., 1990, J. Invertebr. Pathol. 56, 175-180). In this work we used hybridization of single-stranded, unidirectional probes to RNA blots to show that the two transcripts, synthesized in vivo in GmDNV-infected Galleria mellonella larvae, are of antiparallel orientation. To determine their coding specificities, polyadenylated RNAs were isolated from hybrids with DNA from the left and right halves of the viral genome and translated in a rabbit reticulocyte system. The 'right,' 2.4-kb hybrid-selected RNA was shown to direct the synthesis of four polypeptides that comigrated with the four viral capsid proteins and were immunoprecipitated with anti-GmDNV serum. Translation of the 'left,' 1.8-kb RNA yielded three polypeptides, none of which was detected among the vital capsid proteins. This type of expression strategy is unique among vertebrate and most invertebrate parvoviruses, which use only one DNA strand to encode all their proteins. On the other hand, the basic organization of parvoviruses, in which the regulatory and structural proteins are encoded, respectively, by two clusters of ORFs located at the left and right halves of the genome, is conserved. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)126-132
Number of pages7
JournalJournal of Invertebrate Pathology
Volume75
Issue number2
DOIs
StatePublished - 1 Jan 2000

Keywords

  • Densovirus
  • Gene expression
  • In vitro translation

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics

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