The pheromone biosynthesis activating neuropeptide (PBAN) receptor of Heliothis virescens: Identification, functional expression, and structure–activity relationships of ligand analogs

Abstract

Pheromone biosynthesis activating neuropeptide (PBAN) promotes synthesis and release of sex pheromones in moths. We have identified and functionally expressed a PBAN receptor from Heliothis virescens (HevPBANR) and elucidated structure-activity relationships of PBAN analogs. Screening of a larval CNS cDNA library revealed three putative receptor subtypes and nucleotide sequence comparisons suggest they are produced through alternative splicing at the 3'-end. RT-PCR amplified preferentially HevPBANR-C from female pheromone glands. CHO cells expressing HevPBANR-C are highly sensitive to PBAN and related analogs, especially those sharing the C-terminal pentapeptide core, FXPRLamide (X = T, S or V). Alanine replacements in the C-terminal hexapeptide (YFTPRLamide) revealed the relative importance of each residue in the active core as follows, R 5 >L 6 >F 2 ≫P 4 >T 3 ≫Y 1 . This study provides a framework for the rational design of PBANR-specific agonists and/or antagonists that could be exploited for disruption of reproductive function in agriculturally important insect pests.

RNA interference of pheromone biosynthesis-activating neuropeptide receptor suppresses mating behavior by inhibiting sex pheromone production in Plutella xylostella (L.)

“…The production of sex pheromone is mainly dependent on an endocrine signal of PBAN, whose molecular event is initiated by binding to PBANr (Matsumoto et al, 2009;Rafaeli, 2009). To date, PBAN receptors have been identified and characterized from various moths (Hull et al, 2004;Rafaeli et al, 2007;Zheng et al, 2007;Kim et al, 2008). The family of PBAN receptors can be classified into two groups depending on the existence of long C-terminus tail: the first PBAN receptor (Hez-PBANr) identified from H. zea induces extracellular Ca 2þ influx and subsequent increase of cAMP level when challenged with PBAN (Choi et al, 2003).…”

“…and S. exigua (Rafaeli et al, 2007;Cheng et al, 2010). Multiple PBAN receptor transcripts (Hev-PBANr-A, -B, and -C) resulting from alternative splicing were identified from H. virescens in larval central nervous system (Kim et al, 2008). Among Hev-PBAN receptors, Hev-PBANr-C was preferentially expressed in the pheromone gland and responsive to PBAN or related peptides in in vitro expression, whereas other two Hev-PBANrs exhibited little or no Ca 2þ response, whose physiological function and molecular pathway in CNS are not elucidated yet.…”

Molecular Mechanisms Underlying Sex Pheromone Production in Moths

“…A striking difference between the two insect PBANRs is the presence of a 67 amino acid C-terminal extension in the B. mori PBANR that contains a motif essential for receptor internalization, suggesting that the B. mori and H. zea PBANRs are regulated by different mechanisms. 35) PBANR homologs have since been identified in the diamondback moth (Plutella xylostella), the Egyptian cotton leafworm (Spodoptera littoralis), 36) the cotton bollworm (Helicoverpa armigera), 37) and the owlet moth (Heliothis virescens), 38) which contains two PBANR isoforms, a short form (342 residues) and a long form (476 residues) that contains the putative internalization motif identified in the B. mori PBANR.…”

Neural control of gene recruitment in metazoans

“…Secretion of pheromones in all the cases studied so far depends on neural mechanisms. In invertebrates, e.g., secretion of pheromones is neurohumorally regulated by neuropeptides such as PBAN (Christensen et al, 1991;Abernathy et al, 1996;Teal et al, 1999;Groot et al, 2005;Nagalakshmi et al, 2007), whose pheromonotropic effect is mediated by a receptor that is produced in the form of three subtypes by alternative splicing (Kim et al, 2008).…”