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Olfactory proteins mediating chemical communication in the navel orangeworm moth, Amyelois transitella.

Leal WS, Ishida Y, Pelletier J, Xu W, Rayo J, Xu X, Ames JB - PLoS ONE (2009)

Bottom Line: We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor.Of these, AtraPBP1 is highly enriched in male antennae.Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH.

View Article: PubMed Central - PubMed

Affiliation: Department of Entomology, University of California Davis, Davis, California, United States of America. wsleal@ucdavis.edu

ABSTRACT

Background: The navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae), is the most serious insect pest of almonds and pistachios in California for which environmentally friendly alternative methods of control--like pheromone-based approaches--are highly desirable. Some constituents of the sex pheromone are unstable and could be replaced with parapheromones, which may be designed on the basis of molecular interaction of pheromones and pheromone-detecting olfactory proteins.

Methodology: By analyzing extracts from olfactory and non-olfactory tissues, we identified putative olfactory proteins, obtained their N-terminal amino acid sequences by Edman degradation, and used degenerate primers to clone the corresponding cDNAs by SMART RACE. Additionally, we used degenerate primers based on conserved sequences of known proteins to fish out other candidate olfactory genes. We expressed the gene encoding a newly identified pheromone-binding protein, which was analyzed by circular dichroism, fluorescence, and nuclear magnetic resonance, and used in a binding assay to assess affinity to pheromone components.

Conclusion: We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor. Of these, AtraPBP1 is highly enriched in male antennae. Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH.

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Phylogenetic relationships of moth PBPs.Four groups were identified (A–D). The dashed line in Group D suggests a possible subdivision into D1 and D2. The following PBPs have been included in phylogenetic analysis: Agrotis ipsilon: AipsPBP1 (AY301985), AipsPBP2 (AY301986); Antheraea pernyi: AperPBP1 (X96773), AperPBP2 (X96860), AperPBP3 (AJ277265); Antheraea polyphemus: ApolPBP1 (X17559), ApolPBP2 (AJ277266), ApolPBP3 (AJ277267); Agrotis segetum: AsegPBP (AF134292); Ascotis selenaria: AselPBP1 (AB285328), AselPBP2 (AB327273); Argyrotaenia velutinana: AvelPBP (AF177641); Bombyx mori: BmorPBP1 (NM_001044029), BmorPBP2 (AM403100), BmorPBP3 (NM_001083626); Choristoneura fumiferana: CfumPBP (AF177642); Choristoneura murinana: CmurPBP (AF177646); Choristoneura parallela: CparPBP (AF177649); Choristoneura pinus: CpinPBP (AF177653); Choristoneura rosaceana: CrosPBP (AF177652); Diaphania indica: DindPBP (AB263115); Epiphyas postvittana: EposPBP1f (AF416587), EposPBP1s (AF416588), EposPBP2 (AF411459), EposPBP3 (EV811597); Helicoverpa armigera: HarmPBP1 (AJ278992), HarmPBP2 (EU647241), HarmPBP3 (AF527054); Helicoverpa assulta: HassPBP1 (AY864775), HassPBP2 (EU316186), HassPBP3 (DQ286414); Heliothis virescens: HvirPBP1 (X96861), HvirPBP2 (AM403491); Heliothis zea: HzeaPBP (AF090191); Lymantria dispar: LdisPBP1 (AF007867), LdisPBP2 (AF007868); Mamestra brassicae: MbraPBP1 (AF051143), MbraPBP2 (AF051142); Mythimna separata: MsepPBP (AB263112); Manduca sexta: MsexPBP1 (AF117593), MsexPBP2 (AF117589), MsexPBP3 (AF117581); Ostrinia furnacalis: OfurPBP (AF133630); Ostrinia nubilalis: OnubPBP (AF133637); Pectinophora gossypiella: PgosPBP (AF177656); Plutella xylostella: PxylPBP1 (FJ201994), PxylPBP2 (AB263118); Samia cynthia ricini: ScytPBP (AB039793); Synanthodon exitiosa: SexitPBP (AF177657); Spodoptera littoralis: SlitPBP1 (EF396284); Spodoptera litura: SlituPBP1 (DQ004497), SlituPBP2 (DQ114219); Sesamia nonagrioides: SnonPBP1 (AY485219), SnonPBP2 (AY485220); Spodoptera exigua: SexiPBP1 (AY743351), SexiPBP2 (AY545636); Yponomeuta cagnagellus: YcagPBP (AF177661).
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pone-0007235-g007: Phylogenetic relationships of moth PBPs.Four groups were identified (A–D). The dashed line in Group D suggests a possible subdivision into D1 and D2. The following PBPs have been included in phylogenetic analysis: Agrotis ipsilon: AipsPBP1 (AY301985), AipsPBP2 (AY301986); Antheraea pernyi: AperPBP1 (X96773), AperPBP2 (X96860), AperPBP3 (AJ277265); Antheraea polyphemus: ApolPBP1 (X17559), ApolPBP2 (AJ277266), ApolPBP3 (AJ277267); Agrotis segetum: AsegPBP (AF134292); Ascotis selenaria: AselPBP1 (AB285328), AselPBP2 (AB327273); Argyrotaenia velutinana: AvelPBP (AF177641); Bombyx mori: BmorPBP1 (NM_001044029), BmorPBP2 (AM403100), BmorPBP3 (NM_001083626); Choristoneura fumiferana: CfumPBP (AF177642); Choristoneura murinana: CmurPBP (AF177646); Choristoneura parallela: CparPBP (AF177649); Choristoneura pinus: CpinPBP (AF177653); Choristoneura rosaceana: CrosPBP (AF177652); Diaphania indica: DindPBP (AB263115); Epiphyas postvittana: EposPBP1f (AF416587), EposPBP1s (AF416588), EposPBP2 (AF411459), EposPBP3 (EV811597); Helicoverpa armigera: HarmPBP1 (AJ278992), HarmPBP2 (EU647241), HarmPBP3 (AF527054); Helicoverpa assulta: HassPBP1 (AY864775), HassPBP2 (EU316186), HassPBP3 (DQ286414); Heliothis virescens: HvirPBP1 (X96861), HvirPBP2 (AM403491); Heliothis zea: HzeaPBP (AF090191); Lymantria dispar: LdisPBP1 (AF007867), LdisPBP2 (AF007868); Mamestra brassicae: MbraPBP1 (AF051143), MbraPBP2 (AF051142); Mythimna separata: MsepPBP (AB263112); Manduca sexta: MsexPBP1 (AF117593), MsexPBP2 (AF117589), MsexPBP3 (AF117581); Ostrinia furnacalis: OfurPBP (AF133630); Ostrinia nubilalis: OnubPBP (AF133637); Pectinophora gossypiella: PgosPBP (AF177656); Plutella xylostella: PxylPBP1 (FJ201994), PxylPBP2 (AB263118); Samia cynthia ricini: ScytPBP (AB039793); Synanthodon exitiosa: SexitPBP (AF177657); Spodoptera littoralis: SlitPBP1 (EF396284); Spodoptera litura: SlituPBP1 (DQ004497), SlituPBP2 (DQ114219); Sesamia nonagrioides: SnonPBP1 (AY485219), SnonPBP2 (AY485220); Spodoptera exigua: SexiPBP1 (AY743351), SexiPBP2 (AY545636); Yponomeuta cagnagellus: YcagPBP (AF177661).

Mentions: In order to gain insight of the relationships among moth PBPs, we have carried out a phylogenetic analysis in Mega v4.0.2 [33], combining amino acid sequences of the two PBPs from the navel orangeworm (this study) with 57 PBPs previously identified in 33 moth species. A consensus sequence comparison tree was constructed by the neighbor joining method [34] with one thousand bootstrap replicates. The resulting tree suggests that based on their amino acid identity, moth PBPs are clustered into different groups, each comprising related proteins of different moth species (Fig. 7). Indeed, phylogenetic analysis shows the existence of at least four distinct groups of PBPs in moths, illustrating the diversity of this multigenic family. AtraPBP1 and AtraPBP2 belong to two separated groups, with the protein enriched in male antennae, AtraPBP1, clustering with some of the most well-characterized insect PBPs like BmorPBP1 [3], [4], [6], [7], [8], [9], [35], [36] and ApolPBP1 [37], [38], [39], [40]. Despite little boostrap support in the tree, these moth PBPs share 65–70% amino acid identity with AtraPBP1, whereas AtraPBP2 is only 48% identical to AtraPBP1. Contrarily to AtraPBP1, AtraPBP2 belongs to a well supported group (95% bootstrap support) comprising 13 PBPs of other moth species.


Olfactory proteins mediating chemical communication in the navel orangeworm moth, Amyelois transitella.

Leal WS, Ishida Y, Pelletier J, Xu W, Rayo J, Xu X, Ames JB - PLoS ONE (2009)

Phylogenetic relationships of moth PBPs.Four groups were identified (A–D). The dashed line in Group D suggests a possible subdivision into D1 and D2. The following PBPs have been included in phylogenetic analysis: Agrotis ipsilon: AipsPBP1 (AY301985), AipsPBP2 (AY301986); Antheraea pernyi: AperPBP1 (X96773), AperPBP2 (X96860), AperPBP3 (AJ277265); Antheraea polyphemus: ApolPBP1 (X17559), ApolPBP2 (AJ277266), ApolPBP3 (AJ277267); Agrotis segetum: AsegPBP (AF134292); Ascotis selenaria: AselPBP1 (AB285328), AselPBP2 (AB327273); Argyrotaenia velutinana: AvelPBP (AF177641); Bombyx mori: BmorPBP1 (NM_001044029), BmorPBP2 (AM403100), BmorPBP3 (NM_001083626); Choristoneura fumiferana: CfumPBP (AF177642); Choristoneura murinana: CmurPBP (AF177646); Choristoneura parallela: CparPBP (AF177649); Choristoneura pinus: CpinPBP (AF177653); Choristoneura rosaceana: CrosPBP (AF177652); Diaphania indica: DindPBP (AB263115); Epiphyas postvittana: EposPBP1f (AF416587), EposPBP1s (AF416588), EposPBP2 (AF411459), EposPBP3 (EV811597); Helicoverpa armigera: HarmPBP1 (AJ278992), HarmPBP2 (EU647241), HarmPBP3 (AF527054); Helicoverpa assulta: HassPBP1 (AY864775), HassPBP2 (EU316186), HassPBP3 (DQ286414); Heliothis virescens: HvirPBP1 (X96861), HvirPBP2 (AM403491); Heliothis zea: HzeaPBP (AF090191); Lymantria dispar: LdisPBP1 (AF007867), LdisPBP2 (AF007868); Mamestra brassicae: MbraPBP1 (AF051143), MbraPBP2 (AF051142); Mythimna separata: MsepPBP (AB263112); Manduca sexta: MsexPBP1 (AF117593), MsexPBP2 (AF117589), MsexPBP3 (AF117581); Ostrinia furnacalis: OfurPBP (AF133630); Ostrinia nubilalis: OnubPBP (AF133637); Pectinophora gossypiella: PgosPBP (AF177656); Plutella xylostella: PxylPBP1 (FJ201994), PxylPBP2 (AB263118); Samia cynthia ricini: ScytPBP (AB039793); Synanthodon exitiosa: SexitPBP (AF177657); Spodoptera littoralis: SlitPBP1 (EF396284); Spodoptera litura: SlituPBP1 (DQ004497), SlituPBP2 (DQ114219); Sesamia nonagrioides: SnonPBP1 (AY485219), SnonPBP2 (AY485220); Spodoptera exigua: SexiPBP1 (AY743351), SexiPBP2 (AY545636); Yponomeuta cagnagellus: YcagPBP (AF177661).
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pone-0007235-g007: Phylogenetic relationships of moth PBPs.Four groups were identified (A–D). The dashed line in Group D suggests a possible subdivision into D1 and D2. The following PBPs have been included in phylogenetic analysis: Agrotis ipsilon: AipsPBP1 (AY301985), AipsPBP2 (AY301986); Antheraea pernyi: AperPBP1 (X96773), AperPBP2 (X96860), AperPBP3 (AJ277265); Antheraea polyphemus: ApolPBP1 (X17559), ApolPBP2 (AJ277266), ApolPBP3 (AJ277267); Agrotis segetum: AsegPBP (AF134292); Ascotis selenaria: AselPBP1 (AB285328), AselPBP2 (AB327273); Argyrotaenia velutinana: AvelPBP (AF177641); Bombyx mori: BmorPBP1 (NM_001044029), BmorPBP2 (AM403100), BmorPBP3 (NM_001083626); Choristoneura fumiferana: CfumPBP (AF177642); Choristoneura murinana: CmurPBP (AF177646); Choristoneura parallela: CparPBP (AF177649); Choristoneura pinus: CpinPBP (AF177653); Choristoneura rosaceana: CrosPBP (AF177652); Diaphania indica: DindPBP (AB263115); Epiphyas postvittana: EposPBP1f (AF416587), EposPBP1s (AF416588), EposPBP2 (AF411459), EposPBP3 (EV811597); Helicoverpa armigera: HarmPBP1 (AJ278992), HarmPBP2 (EU647241), HarmPBP3 (AF527054); Helicoverpa assulta: HassPBP1 (AY864775), HassPBP2 (EU316186), HassPBP3 (DQ286414); Heliothis virescens: HvirPBP1 (X96861), HvirPBP2 (AM403491); Heliothis zea: HzeaPBP (AF090191); Lymantria dispar: LdisPBP1 (AF007867), LdisPBP2 (AF007868); Mamestra brassicae: MbraPBP1 (AF051143), MbraPBP2 (AF051142); Mythimna separata: MsepPBP (AB263112); Manduca sexta: MsexPBP1 (AF117593), MsexPBP2 (AF117589), MsexPBP3 (AF117581); Ostrinia furnacalis: OfurPBP (AF133630); Ostrinia nubilalis: OnubPBP (AF133637); Pectinophora gossypiella: PgosPBP (AF177656); Plutella xylostella: PxylPBP1 (FJ201994), PxylPBP2 (AB263118); Samia cynthia ricini: ScytPBP (AB039793); Synanthodon exitiosa: SexitPBP (AF177657); Spodoptera littoralis: SlitPBP1 (EF396284); Spodoptera litura: SlituPBP1 (DQ004497), SlituPBP2 (DQ114219); Sesamia nonagrioides: SnonPBP1 (AY485219), SnonPBP2 (AY485220); Spodoptera exigua: SexiPBP1 (AY743351), SexiPBP2 (AY545636); Yponomeuta cagnagellus: YcagPBP (AF177661).
Mentions: In order to gain insight of the relationships among moth PBPs, we have carried out a phylogenetic analysis in Mega v4.0.2 [33], combining amino acid sequences of the two PBPs from the navel orangeworm (this study) with 57 PBPs previously identified in 33 moth species. A consensus sequence comparison tree was constructed by the neighbor joining method [34] with one thousand bootstrap replicates. The resulting tree suggests that based on their amino acid identity, moth PBPs are clustered into different groups, each comprising related proteins of different moth species (Fig. 7). Indeed, phylogenetic analysis shows the existence of at least four distinct groups of PBPs in moths, illustrating the diversity of this multigenic family. AtraPBP1 and AtraPBP2 belong to two separated groups, with the protein enriched in male antennae, AtraPBP1, clustering with some of the most well-characterized insect PBPs like BmorPBP1 [3], [4], [6], [7], [8], [9], [35], [36] and ApolPBP1 [37], [38], [39], [40]. Despite little boostrap support in the tree, these moth PBPs share 65–70% amino acid identity with AtraPBP1, whereas AtraPBP2 is only 48% identical to AtraPBP1. Contrarily to AtraPBP1, AtraPBP2 belongs to a well supported group (95% bootstrap support) comprising 13 PBPs of other moth species.

Bottom Line: We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor.Of these, AtraPBP1 is highly enriched in male antennae.Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH.

View Article: PubMed Central - PubMed

Affiliation: Department of Entomology, University of California Davis, Davis, California, United States of America. wsleal@ucdavis.edu

ABSTRACT

Background: The navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae), is the most serious insect pest of almonds and pistachios in California for which environmentally friendly alternative methods of control--like pheromone-based approaches--are highly desirable. Some constituents of the sex pheromone are unstable and could be replaced with parapheromones, which may be designed on the basis of molecular interaction of pheromones and pheromone-detecting olfactory proteins.

Methodology: By analyzing extracts from olfactory and non-olfactory tissues, we identified putative olfactory proteins, obtained their N-terminal amino acid sequences by Edman degradation, and used degenerate primers to clone the corresponding cDNAs by SMART RACE. Additionally, we used degenerate primers based on conserved sequences of known proteins to fish out other candidate olfactory genes. We expressed the gene encoding a newly identified pheromone-binding protein, which was analyzed by circular dichroism, fluorescence, and nuclear magnetic resonance, and used in a binding assay to assess affinity to pheromone components.

Conclusion: We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor. Of these, AtraPBP1 is highly enriched in male antennae. Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH.

Show MeSH
Related in: MedlinePlus