title: Identification of an in vitro interaction between an insect immune suppressor protein (CrV2) and G alpha proteins.
abstract: The protein CrV2 is encoded by a polydnavirus integrated into the genome of the endoparasitoid Cotesia rubecula (Hymenoptera:Braconidae:Microgastrinae) and is expressed in host larvae with other gene products of the polydnavirus to allow successful development of the parasitoid. CrV2 expression has previously been associated with immune suppression, although the molecular basis for this was not known. Here, we have used time-resolved Förster resonance energy transfer (TR-FRET) to demonstrate high affinity binding of CrV2 to Gα subunits (but not the Gβγ dimer) of heterotrimeric G-proteins. Signals up to 5-fold above background were generated, and an apparent dissociation constant of 6.2 nm was calculated. Protease treatment abolished the TR-FRET signal, and the presence of unlabeled CrV2 or Gα proteins also reduced the TR-FRET signal. The activation state of the Gα subunit was altered with aluminum fluoride, and this decreased the affinity of the interaction with CrV2. It was also demonstrated that CrV2 preferentially bound to Drosophila Gα(o) compared with rat Gα(i1). In addition, three CrV2 homologs were detected in sequences derived from polydnaviruses from Cotesia plutellae and Cotesia congregata (including the immune-related early expressed transcript, EP2). These data suggest a potential mode-of-action of immune suppressors not previously reported, which in addition to furthering our understanding of insect immunity may have practical benefits such as facilitating development of novel controls for pest insect species.
authors.affiliation: South Australian Research and Development Institute, Entomology, Waite Road, Urrbrae, South Australia 5064, Australia.
authors.forename: Tamara H
authors.initials: TH
authors.lastname: Cooper
authors.forename: Kelly
authors.initials: K
authors.lastname: Bailey-Hill
authors.forename: Wayne R
authors.initials: WR
authors.lastname: Leifert
authors.forename: Edward J
authors.initials: EJ
authors.lastname: McMurchie
authors.forename: Sassan
authors.initials: S
authors.lastname: Asgari
authors.forename: Richard V
authors.initials: RV
authors.lastname: Glatz
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral / Q000276:immunology*; D007108:Immune Tolerance / Q000276:immunology*; D019476:Insect Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018071:Polydnaviridae / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014863:Wasps / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000821:virology
chemical_list: D044385:GTP-Binding Protein alpha Subunits; D019476:Insect Proteins; D014764:Viral Proteins