title: Prophenoloxidases 1 and 2 from the spruce budworm, Choristoneura fumiferana: molecular cloning and assessment of transcriptional regulation by a polydnavirus.
abstract: Immune challenge in arthropods is frequently accompanied by melanization of the hemolymph, a reaction triggered by the activation of prophenoloxidase (PPO). Because their immature stages are spent inside the hemocoel of insect larvae, endoparasitoids have evolved strategies to escape or counter melanin formation. Very little molecular information is available on these endoparasitoid counterstrategies. We have sought to shed light on the inhibition of melanization in the spruce budworm, Choristoneura fumiferana, by the parasitic wasp Tranosema rostrale, by cloning two host PPO homologs and studying their transcriptional regulation after parasitization. The two polypeptides are encoded by transcripts of approximately 3.3 kb (for CfPPO1) and 3.0 kb (for CfPPO2) and possess structural features typical of other insect PPOs. While there appears to be a single CfPPO2 gene in the C. fumiferana genome, we detected three CfPPO1 mRNA variants displaying insertions/deletions in the 3' untranslated region, suggesting that there may be more than one CfPPO1 gene copy. Both CfPPO1 and CfPPO2 were expressed at high levels in C. fumiferana 6th instars, and parasitization by T. rostrale had no apparent impact on the level of their transcripts. Injection of a large dose (0.5 female-equivalent) of polydnavirus-laden calyx fluid extracted from T. rostrale, which is known to inhibit melanization in C. fumiferana, only caused a transient decrease in CfPPO1 and CfPPO2 transcript accumulation at 2-3 d post injection. It thus appears that transcriptional downregulation of C. fumiferana PPO by T. rostrale plays a minor role in the inhibition of hemolymph melanization in this host-parasitoid system.
authors.affiliation: Laurentian Forestry Centre, Natural Resources Canada, Quebec, Canada. Dan.Doucet@NRCAN.gc.ca
authors.forename: Daniel
authors.initials: D
authors.lastname: Doucet
authors.forename: Catherine
authors.initials: C
authors.lastname: Béliveau
authors.forename: Ashley
authors.initials: A
authors.lastname: Dowling
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Simard
authors.forename: Qili
authors.initials: Q
authors.lastname: Feng
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Krell
authors.forename: Michel
authors.initials: M
authors.lastname: Cusson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D004156:Catechol Oxidase / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics*; D040461:Gene Components; D005786:Gene Expression Regulation / Q000276:immunology*; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology* / Q000276:immunology / Q000469:parasitology*; D010802:Phylogeny; D018071:Polydnaviridae; D017422:Sequence Analysis, DNA; D014863:Wasps / Q000276:immunology*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004792:Enzyme Precursors; C038451:pro-phenoloxidase; D004156:Catechol Oxidase