title: Functional annotation of Cotesia congregata bracovirus: identification of viral genes expressed in parasitized host immune tissues.
abstract: Bracoviruses (BVs) from the Polydnaviridae family are symbiotic viruses used as biological weapons by parasitoid wasps to manipulate lepidopteran host physiology and induce parasitism success. BV particles are produced by wasp ovaries and injected along with the eggs into the caterpillar host body, where viral gene expression is necessary for wasp development. Recent sequencing of the proviral genome of Cotesia congregata BV (CcBV) identified 222 predicted virulence genes present on 35 proviral segments integrated into the wasp genome. To date, the expressions of only a few selected candidate virulence genes have been studied in the caterpillar host, and we lacked a global vision of viral gene expression. In this study, a large-scale transcriptomic analysis by 454 sequencing of two immune tissues (fat body and hemocytes) of parasitized Manduca sexta caterpillar hosts allowed the detection of expression of 88 CcBV genes expressed 24 h after the onset of parasitism. We linked the expression profiles of these genes to several factors, showing that different regulatory mechanisms control viral gene expression in the host. These factors include the presence of signal peptides in encoded proteins, diversification of promoter regions, and, more surprisingly, gene position on the proviral genome. Indeed, most genes for which expression could be detected are localized in particular proviral regions globally producing higher numbers of circles. Moreover, this polydnavirus (PDV) transcriptomic analysis also reveals that a majority of CcBV genes possess at least one intron and an arthropod transcription start site, consistent with an insect origin of these virulence genes.
  
  
  OBJECTIVE
  Bracoviruses (BVs) are symbiotic polydnaviruses used by parasitoid wasps to manipulate lepidopteran host physiology, ensuring wasp offspring survival. To date, the expressions of only a few selected candidate BV virulence genes have been studied in caterpillar hosts. We performed a large-scale analysis of BV gene expression in two immune tissues of Manduca sexta caterpillars parasitized by Cotesia congregata wasps. Genes for which expression could be detected corresponded to genes localized in particular regions of the viral genome globally producing higher numbers of circles. Our study thus brings an original global vision of viral gene expression and paves the way to the determination of the regulatory mechanisms enabling the expression of BV genes in targeted organisms, such as major insect pests. In addition, we identify sequence features suggesting that most BV virulence genes were acquired from insect genomes.
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France germain.chevignon@etu.univ-tours.fr elisabeth.huguet@univ-tours.fr.
authors.forename: Germain
authors.initials: G
authors.lastname: Chevignon
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Thézé
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Cambier
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives, Genoscope (Centre National de Séquençage), Evry, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Poulain
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives, Genoscope (Centre National de Séquençage), Evry, France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Da Silva
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Annie
authors.initials: A
authors.lastname: Bézier
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Musset
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Sébastien J M
authors.initials: SJ
authors.lastname: Moreau
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Drezen
authors.affiliation: Institut de Recherche sur la Biologie de l'Insecte, UMR CNRS 7261, UFR Sciences et Techniques, Université François-Rabelais, Tours, France germain.chevignon@etu.univ-tours.fr elisabeth.huguet@univ-tours.fr.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Huguet
mesh_terms: D000818:Animals; D015870:Gene Expression / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D005814:Genes, Viral / Q000235:genetics*; D016679:Genome, Viral / Q000235:genetics*; D018402:Manduca / Q000235:genetics / Q000821:virology; D018071:Polydnaviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014863:Wasps / Q000235:genetics* / Q000821:virology*