The pheromone biosynthesis activating neuropeptide (PBAN) of the black cutworm moth, Agrotis ipsilon: immunohistochemistry, molecular characterization and bioassay of its peptide sequence.
PBAN-like immunoreactivity has been detected in the suboesophageal ganglion and the brain (Br-SOG) of larvae and adult males and females of Agrotis ipsilon, using an antiserum against Helicoverpa zea PBAN (Hez-PBAN). The amino acid sequence of A. ipsilon PBAN (Agi-PBAN) was deduced from the cDNA sequence, using both Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and 5' Rapid Amplification of cDNA Ends (RACE). The primers were degenerate sets of oligonucleotides derived from known amino acid sequences of the PBAN precursor. The final cloned fragment contained the complete DNA sequence coding for the putative Agi-PBAN. Based on a comparison with known PBAN processing from the polypeptide precursor, we propose that Agi-PBAN is a 33-amino acid peptide. Agi-PBAN exhibits high sequence homology with Hez-PBAN (88%), Lymantria dispar PBAN (Lyd-PBAN, 88%) and Bombyx mori PBAN (Bom-PBAN, 73%). Agi-PBAN shares the C-terminal hexapeptide sequence (Tyr-Phe-Ser-Pro-Arg-LeuNH2) with all identified PBANs but has only one methionine residue instead of two in Hez-PBAN and Lyd-PBAN, and three in Bom-PBAN. Based on predicted a.a. sequence, Agi-PBAN, with Leu-NH2 as C-terminal motif, has been synthesized and assayed for its ability to promote pheromone production in decapitated females of A. ipsilon. Synthetic Agi-PBAN induced pheromone production in decapitated females as evaluated by the male responsiveness to the pheromonal blend in a wind tunnel.
Duportets
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Laboratoire de Neuroendocrinologie, Université Bordeaux I, ER 629, Talence, France.
Gadenne
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Dufour
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MC
Couillaud
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D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D007814:Larva / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics* / Q000502:physiology; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012724:Sex Attractants / Q000096:biosynthesis* / Q000235:genetics
D018076:DNA, Complementary; D009479:Neuropeptides; D012724:Sex Attractants